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  • Articles  (118)
  • gene expression  (65)
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  • Springer  (118)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular life sciences 51 (1995), S. 1116-1123 
    ISSN: 1420-9071
    Keywords: Antisense RNA ; gene expression ; insertional mutagenesis ; physical mapping ; reporter genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Over the past ten years, powerful molecular genetic techniques have been developed to analyze gene function inDictyostelium. DNA-mediated transformation using a variety of selections and vectors has allowed the introduction of wild-type or modified genes that are under various forms of transcriptional control. Homologous recombination is efficient and can be used to modify the genome in precise ways. In addition, it is now possible to clone genes based on their mutant phenotype alone, either by insertional mutagenesis, or by screening antisense expression cDNA libraries. Finally, a nearly complete physical map of the genome is available and so genes are easily mapped by physical techniques. We discuss many of these advances within the context of major research problems presently under study.
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  • 2
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    Cellular and molecular life sciences 51 (1995), S. 606-611 
    ISSN: 1420-9071
    Keywords: Metallothionein ; isometallothioneins ; gene expression ; rabbit kidney cell-line ; cadmium adaptation ; zinc adaptation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We explored the molecular genetics underlying the massive induction of isoMTs by Zn2+ or Cd2+ in metal tolerant rabbit kidney (RK-13) sub-line cells, using band shift assays and Southern blotting analysis. In sub-line cells accommodated to intermediate metal concentrations (100 μM Zn2+; 1–20 μM Cd2+) evidence suggested that the increase in the capacity for isoMT synthesis is brought about by an increased binding activity of the nuclear transcription factors MTF-1 and Sp1. Using quantitative band shift analysis with a mouse MRE-d oligonucleotide probe, the binding of both transcription factors was found to be enhanced two to three times over the binding activity measured in the unexposed parental RK-13 cells. Their increase in binding activity is probably the cause of the overexpression of MT genes and the development of metal tolerance in these cells. In cells tolerant to the highest concentrations of metal the analysis of Southern blot signals revealed MT gene amplification to be the most probable cause of the increased MT production. Thus, in cells of sub-lines growing in the presence of 350 μM Zn2+, two of the isoMT genes were coordinately triplicated and in cells tolerant to 150 μM Cd2+ one isoMT gene was amplified two-fold.
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  • 3
    ISSN: 1432-1432
    Keywords: Evolution ; κ-Casein ; Cytochrome b ; Artiodactyla ; Ruminantia ; Caprinae ; Capricornis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nucleotide sequences for the κ-casein precursor proteins have been determined from the genomic DNAs or hair roots of the Ruminantia. The coding regions, exons 2, 3, and 4, were amplified separately via the three kinds of PCRs and then directly sequenced. The primers were designed from the sequence of bovine κ-casein gene; they were applicable for the amplification of the κ-casein genes from the 13 species in the Ruminantia except exon 2 of the lesser mouse deer. These results permitted an easy phylogenetic analysis based on the sequences of an autosomal gene. A phylogenetic tree was constructed from the mature K-casein sequences and compared with the tree of the cytochrome b genes which were sequenced from the same individuals. The Cervidae (sika deer, Cervus nippon) were separated from the branch of the Bovidae on the tree of κ-casein genes with a relatively high confidence level of the bootstrap analysis, but included in the branch of the Bovidae on the tree of cytochrome b genes. The κ-casein tree indicated a monophyly of the subfamily Caprinae, although the internal branches were uncertain in the Caprinae. The tree based on the nucleotide sequences of cytochrome b genes clearly showed the relationships of the closely related species in the genus Capricornis consisting of serow (C. smatorensis), Japanese serow (C. crispus), and Formosan serow (C. swinhoei). These results would be explained by the difference of resolving power between the κ-casein and the cytochrome b sequences.
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  • 4
    ISSN: 1432-1432
    Keywords: Evolution ; Expression patterns ; α-Amylase ; Glucose repression ; Starch induction ; Intra- and interspecific variation ; Drosophila ; Gene expression ; Regulatory genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Intraspecific variation of α-amylase activity in D. melanogaster and D. immigrans, which is distantly related to D. melanogaster, and interspecific variation of α-amylase activity in 18 Drosophila species were examined. The amount of intraspecific variation of α-amylase activities measured in terms of coefficient of variation in D. melanogaster and D. immigrans was one-half and one-tenth or less, respectively, of the interspecific variation in 18 Drosophila species. We also surveyed the response patterns of α-amylase activity to dietary carbohydrates at the larval and adult stages. The levels of α-amylase activity depended on both repression by dietary glucose (glucose repression) and induction by dietary starch (starch induction). In general, our data suggest that glucose repression was conserved among species at both stages while starch induction was mainly observed in larvae, although the degree of the response depended on species. In D. lebanonensis lebanonensis and D. serrata, larvae expressed electrophoretically different α-amylase variants (isozymes) from those of adult flies. These results may suggest that the regulatory systems responsible both for the response to environment and developmental expression are different among species in Drosophila.
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  • 5
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    Journal of molecular evolution 40 (1995), S. 570-577 
    ISSN: 1432-1432
    Keywords: Alternate reading frames ; Evolution ; Overlapping frames ; Homology search ; Primary sequence analysis ; Polycistronic genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hypothetical Products from Noncoding Frames (i.e., HyPNoFs) are hypothetical, not-coded proteins, translated from alternate reading frames (i.e., coding+1 and coding+2) of cDNAs. HyPNoFs of CD4, PKC, oncostatin, bcl-2 proto-oncogene, tumor suppressor p53, cystic fibrosis transmembrane regulator (CFTR), and tumor necrosis factors a and β were searched as query sequences vs the SWISS-PROT data bank. Homology searches carried out revealed that hypothetical products (i.e., HyPNoFs) may share high similarity with real protein products actually coded. Sequence similarity of hypothetical products to real proteins is sometimes very high, suggesting common conformational features, according to the Sander and Schneider cutoff value. This finding supports the hypothesis that eukaryotic DNA, currently considered to be monocistronic, might occasionally have polycistronic regions, carrying different protein messages on overlapping frames. As yet, polycistronic genes have been observed in viral genomes only. The presence of polycistronic regions in eukaryotic genes is likely reminiscent of an ancient strategy, rather than a present feature of the genome in eukaryotes. These data suggest that thorough investigation of HyPNoFs is likely to improve our ability to trace genes' evolution and to investigate structure-function relationships of protein and DNA sequences.
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  • 6
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    Journal of molecular evolution 40 (1995), S. 601-607 
    ISSN: 1432-1432
    Keywords: Prolamine ; Sperm proteins ; Evolution ; Mammals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Prolamine P1 genes have been sequenced following PCR amplification from 11 mammals representing five major mammalian orders: Rodentia (rat and guinea pig), Carnivora (cat and bear), Proboscidea (elephant), Perissodactyla (horse), and Artiodactyla (camel, deer, elk, moose, and gazelle). The predicted amino acid sequence for these genes together with previously reported sequences results in a data set of 25 different P1 genes and 30 different P1 amino acid sequences. The alignment of all these sequences reveals that prolamines are amongst the most rapidly diverging proteins studied. In spite of the large number of differences there are conserved motifs that are also common to birds such as the N-terminal ARYR followed by the triple alternating SRSRSR phosphorylation site. The central region contains 3 arginine clusters consisting of 5–6 arginines each. The C-terminus appears to be the most variable region of the protamines. Overall the molecular evolution of P1 genes is in agreement with the expected species evolution supporting that these genes have evolved vertically.
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  • 7
    ISSN: 1432-1432
    Keywords: Snake venom ; Bothrops ; Phospholipase ; Myotoxin ; Evolution ; cDNA ; Gene duplication ; Phylogeny
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The sequence coding for a snake venom phospholipase A2 (PLA2), BJUPLA2, has been cloned from a Bothrops jararacussu venom gland cDNA library. The cDNA sequence predicts a precursor containing a 16-residue signal peptide followed by a molecule of 122 amino acid residues with a strong sequence similarity to group II snake venom PLA2's. A striking feature of the cDNA is the high sequence conservation of the 5′ and 3′ untranslated regions in cDNAs coding for PLA2's from a number of viper species. The greatest sequence variation was observed between the regions coding for the mature proteins, with most substitutions occurring in nonsynonymous sites. The phylogenetic tree constructed by alignment of the amino acid sequence of BJUPLA2 with group II PLA2's in general groups them according to current taxonomical divisions and/or functional activity. It also suggests that gene duplications may have occurred at a number of different points during the evolution of snake venom group II PLA2's.
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  • 8
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    Journal of molecular evolution 41 (1995), S. 238-246 
    ISSN: 1432-1432
    Keywords: Cellular slime molds ; Animals ; Fungi ; Plantae ; Maximum-likelihood method ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The phylogenetic position of Dictyostelium inferred from 18S rRNA data contradicts that from protein data. Protein trees always show the close affinity of Dictyostelium with animals, fungi, and plants, whereas in 18S rRNA trees the branching of Dictyostelium is placed at a position before the massive radiation of protist groups including the divergence of the three kingdoms. To settle this controversial issue and to determine the correct position of Dictyostelium, we inferred the phylogenetic relationship among Dictyostelium and the three kingdoms Animalia, Fungi, and Plantae by a maximum-likelihood method using 19 different protein data sets. It was shown at the significance level of 1 SE that the branching of Dictyostelium antedates the divergence of Animalia and Fungi, and Plantae is an outgroup of the Animalia-Fungi-Dictyostelium clade.
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  • 9
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    Journal of molecular evolution 41 (1995), S. 293-298 
    ISSN: 1432-1432
    Keywords: β-tubulin ; Evolution ; Gene cluster ; Gene dispersion ; Drosophila montium subgroup
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The β1-, β2-, and β3-tubulin genes have been mapped by in situ hybridization on the polytene chromosomes of 11 selected species (15 strains) belonging to theDrosophila montium subgroup. Although the hybridization pattern among the strains of the same species does not differ, this pattern is significantly different among the species. The β-tubulin genes in themontium subgroup seem to be organized in a cluster, or in a semi-cluster, or are completely dispersed. The clustered arrangement is found in the North-Oriental sibling speciesD. auraria, D. triauraria, andD. quadraria. The semi-clustered arrangement, wherein the β1 and β2 genes are located at the same locus while β3 is at a different one, appears in the South-Oriental speciesD. bicomuta, D. serrata, andD. birchii, as well as in the Afrotropical speciesD. diplacantha andD. seguyi. The complete separation of the genes is observed in the Indian speciesD. kikkawai andD. jambulina and in the Afrotropical speciesD. vulcana. Based on the above results, a possible mode of evolution of the β-tubulin genes in the montium subgroup is attempted. In addition, phylogenetic relationships among themontium species are discussed.
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  • 10
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    Journal of molecular evolution 41 (1995), S. 293-298 
    ISSN: 1432-1432
    Keywords: β-tubulin ; Evolution ; Gene cluster ; Gene dispersion ; Drosophila montium subgroup
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The β1-, β2-, and β3-tubulin genes have been mapped by in situ hybridization on the polytene chromosomes of 11 selected species (15 strains) belonging to the Drosophila montium subgroup. Although the hybridization pattern among the strains of the same species does not differ, this pattern is significantly different among the species. The β-tubulin genes in the montium subgroup seem to be organized in a cluster, or in a semi-cluster, or are completely dispersed. The clustered arrangement is found in the North-Oriental sibling species D. auraria, D. triauraria, and D. quadraria. The semi-clustered arrangement, wherein the β1 and β2 genes are located at the same locus while β3 is at a different one, appears in the South-Oriental species D. bicomuta, D. serrata, and D. birchii, as well as in the Afrotropical species D. diplacantha and D. seguyi. The complete separation of the genes is observed in the Indian species D. kikkawai and D. jambulina and in the Afrotropical species D. vulcana. Based on the above results, a possible mode of evolution of the β-tubulin genes in the montium subgroup is attempted. In addition, phylogenetic relationships among the montium species are discussed.
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  • 11
    ISSN: 1432-1432
    Keywords: Giardia ; Trichomonas ; CDK ; CDC ; Unicellular ; Metazoa ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyclin-dependent kinase (CDK) and cell division control (CDC2) sequences are strongly conserved among eukaryotes and may complement the use of other sequence families in eukaryotic phylogenetic inference. We synthesized degenerate PCR primers to amplify the catalytic region of CDK homologs in representatives of the earliest available lineages of eukaryotes. CDK family sequence-based, maximum-likelihood distance measurements with neighbor joining, and Fitch-Margoliash least-squares analyses produced unrooted dendrograms that included protists, yeasts, and higher eukaryotes. Bootstrap confidence estimates supported CDK-based phylogenetic groupings among the protists, fungi, and vertebrates although resolution within these groups was often insignificant. However, Trichomonas vaginalis and Giardia lamblia exhibited two of the most divergent CDK-like sequences consistent with rRNA-phylogenetic inference of early divergence of these eukaryotic lineages. In the evolution from unicellular to multicellular organisms, a constellation of amino acid residues aligning with the human, CDK N-terminal β sheet may have undergone abrupt replacement.
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  • 12
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    Journal of molecular evolution 41 (1995), S. 813-832 
    ISSN: 1432-1432
    Keywords: Phylogeny ; Evolution ; Carbamoylphosphate synthetase ; Dihydroorotase ; Aspartate transcarbamoylase ; Intron ; Exon ; Arginine biosynthesis ; Pyrimidine biosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Carbamoylphosphate is a common intermediate in the metabolic pathways leading to the biosynthesis of arginine and pyrimidines. The amino acid sequences of all available proteins that catalyze the formation of carbamoylphosphate were retrieved from Genbank and aligned to estimate their mutual phylogenetic relations. In gram-negative bacteria carbamoylphosphate is synthesized by a two-subunit enzyme with glutamiriase and carbamoylphosphate synthetase (CPS) activity, respectively. In gram-positive bacteria and lower eukaryotes this two-subunit CPS has become dedicated to arginine biosynthesis, while in higher eukaryotes the two subunits fused and subsequently lost the glutaminase activity. The CPS dedicated to pyrimidine synthesis is part of a multifunctional enzyme (CPS II), encoding in addition dihydroorotase and aspartate transcarbamoylase. Evidence is presented to strengthen the hypothesis that the two “kinas” subdomains of all CPS isozymes arose from a duplication of an ancestral gene in the progenote. A further duplication of the entire CPS gene occurred after the divergence of the plants and before the divergence of the fungi from the eukaryotec root, generating the two isoenzymes involved in either the synthesis of arginine or that of pyrimidines. The mutation rate was found to be five- to tenfold higher after the duplication than before, probably reflecting optimization of the enzymes for their newly acquired specialized function. We hypothesize that this duplication arose from a need for metabolic channeling for pyrimidine biosynthesis as it was accompanied by the tagging of the CPS gene with the genes for dihydroorotase and aspartate transcarbamoylase, and as the duplication occurred independently also in gram-positive bacteria. Analysis of the exon-intron organization of the two “kinase” subdomains in CPS I and II suggests that ancient exons may have comprised approx. 19 amino acids, in accordance with the prediction of the “intron-early” theory.
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  • 13
    ISSN: 1432-1432
    Keywords: Ribonuclease ; Gene duplication ; Evolution ; Ruminants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phylogenetic analysis, based on the primary structures of mammalian pancreatic-type ribonucleases, indicated that gene duplication events, which occurred during the evolution of ancestral ruminants, gave rise to the three paralogous enzymes present in the bovine species. Herein we report data that demonstrate the existence of the orthologues of the bovine pancreatic, seminal, and cerebral ribonucleases coding sequences in the genomes of giraffe and sheep. The “seminal” sequence is a pseudogene in both species. We also report an analysis of the transcriptional expression of ribonuclease genes in sheep tissues. The data presented support a model for positive selection acting on the molecular evolution of ruminant ribonuclease genes.
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  • 14
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    Journal of molecular evolution 40 (1995), S. 482-486 
    ISSN: 1432-1432
    Keywords: Aminoacyl-tRNA synthetase ; Computer analysis ; Evolution ; Genetic code ; tRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract tRNA sequences were analyzed for sequence features correlated with known classes of aminoacyl-tRNA synthetase enzymes. The tRNAs were searched for distinguishing nucleotides anywhere in their sequences. The analyses did not find nucleotides predictive of synthetase class membership. We conclude that such nucleotides never existed in tRNA sequences or that they existed and were lost from many of the tRNA sequences during evolution.
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  • 15
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    Journal of molecular evolution 40 (1995), S. 476-481 
    ISSN: 1432-1432
    Keywords: Aminoacylation ; Aminoacyl-tRNA synthetases ; tRNA ; GluRS ; GlnRS ; Glutamate ; Glutamine ; Evolution ; Mischarging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Aminoacyl-tRNA for protein synthesis is produced through the action of a family of enzymes called aminoacyl-tRNA synthetases. A general rule is that there is one aminoacyl-tRNA synthetase for each of the standard 20 amino acids found in all cells. This is not universal, however, as a majority of prokaryotic organisms and eukaryotic organelles lack the enzyme glutaminyl-tRNA synthetase, which is responsible for forming Gln-tRNAGln in eukaryotes and in Gram-negative eubacteria. Instead, in organisms lacking glutaminyl-tRNA synthetase, Gln-tRNAGln is provided by misacylation of tRNAGln with glutamate by glutamyl-tRNA synthetase, followed by the conversion of tRNA-bound glutamate to glutamine by the enzyme Glu-tRNAGln amidotransferase. The fact that two different pathways exist for charging glutamine tRNA indicates that ancestral prokaryotic and eukaryotic organisms evolved different cellular mechanisms for incorporating glutamine into proteins. Here, we explore the basis for diverging pathways for aminoacylation of glutamine tRNA. We propose that stable retention of glutaminyl-tRNA synthetase in prokaryotic organisms following a horizontal gene transfer event from eukaryotic organisms (Lamour et al. 1994) was dependent on the evolving pool of glutamate and glutamine tRNAs in the organisms that acquired glutaminyl-tRNA synthetase by this mechanism. This model also addresses several unusual aspects of aminoacylation by glutamyl- and glutaminyl-tRNA synthetases that have been observed.
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  • 16
    ISSN: 1432-1432
    Keywords: Aminoacyl-tRNA synthetases ; tRNA ; Genetic code ; RNA world ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two aspects of the evolution of aminoacyl-tRNA synthetases are discussed. Firstly, using recent crystal structure information on seryl-tRNA synthetase and its substrate complexes, the coevolution of the mode of recognition between seryl-tRNA synthetase and tRNAser in different organisms is reviewed. Secondly, using sequence alignments and phylogenetic trees, the early evolution of class 2 Amnoacyl-tRNA synthetases is traced. Arguments are presented to suggest that synthetases are not the oldest of protein enzymes, but survived as RNA enzymes during the early period of the evolution of protein catalysts. In this view, the relatedness of the current synthetases, as evidenced by the division into two classes with their associated subclasses, reflects the replacement of RNA synthetases by protein synthetases. This process would have been triggered by the acquisition of tRNA 3′ end charging activity by early proteins capable of activating small molecules (e.g., amino acids) with ATP. If these arguments are correct, the genetic code was essentially frozen before the protein synthetases that we know today came into existence. Correspondence to: S. Cusack
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  • 17
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    Journal of molecular evolution 41 (1995), S. 996-1005 
    ISSN: 1432-1432
    Keywords: Repetitive DNA ; Tandem repeats ; Sequence analysis ; Phylogenetic tree ; Silent repeats ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two different satellite DNAs exist in the genus Cucurbita which are different with respect to repeat length (350 by and 170 bp), array size, and sequence homogenization. Whereas the 350-bp satellite DNA is prominent and very homogeneous in all species investigated except for C. maxima and C. lundelliana, the 170-bp satellite is rather evenly distributed in all species. In C. maxima and C. lundelliana the 350-bp satellite is present only in small amounts, but detectable by the sensitive PCR method. These repeats are also very homogeneous, reflecting a silent stage of satellite DNA. In contrast, the 170-bp satellite DNA is intra- and interspecifically heterogeneous. It is striking that the species with no detectable amount of 350-bp satellite contain 170-bp satellite DNA clusters with the highest degree of homogeneity. The evolution of satellite DNA repeats within cultivated and wild species in the genus Cucurbita is elucidated using the sequence data of both satellite DNAs from all species investigated. The value of satellite DNA for phylogenetic analysis between closely related species is discussed.
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  • 18
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    Journal of molecular evolution 40 (1995), S. 120-126 
    ISSN: 1432-1432
    Keywords: Simple repetitive DNA ; Microsatellites ; Unstable triplets ; Database ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Simple repeats composed of tandemly repeated units 1–6 nucleotides (nt) long have been extracted from a selected set of primate genomic DNA sequences. Of the 501 theoretically possible, different types of repeats only 67 were present in the analyzed database in at least two different size ranges over 12 nt. They include all simple repeats known to be polymorphic in the primate genome. A list of moderately expanding and nonexpanding oligonucleotide patterns has also been included. Furthermore, we have compiled statistical data with emphasis on the overall variability of the most abundant 67 types of repeats. We have demonstrated that the expandability of at least some simple repeats may be affected by the overall base composition and by flanking sequences. In particular, the occurrence of tandemly repeated CAG and GCC triplets in exons positively correlates with their G+C content. We also noted that in the vicinity of Alu sequences tetrameric repeats are more abundant than in the total genomic DNA. This paper can be used as a comprehensive guide in identification of the most abundant and potentially polymorphic simple repeats. It is also of broader significance as a step toward understanding the contribution of flanking sequences and the overall sequence composition to variability of simple repeats.
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  • 19
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    Journal of molecular evolution 41 (1995), S. 457-466 
    ISSN: 1432-1432
    Keywords: Seed storage proteins ; Legumin ; Gene structure ; Evolution ; Gymnosperms ; Ginkgo biloba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Legumin-like seed storage proteins have been intensively studied in crop plants. However, little is known about the molecular evolution of these proteins and their genes and it was assumed that they originated from an ancestral gene that already existed at the beginning of angiosperm evolution. We have evidence for the ubiquitous occurrence of homologous proteins in gymnosperms as well. We have characterized the major seed storage globulin from Ginkgo biloba by amino acid sequencing, which reveals clear homology to legumin-like proteins from angiosperms. The Ginkgo legumin is encoded by a gene family; we describe two of its members. The promoter regions contain sequence motifs which are known to function as regulatory elements involved in seed-specific expression of angiosperm legumins, although the tissues concerned are different in gymnosperms and angiosperms. The Ginkgo legumin gene structure is divergent from that of angiosperms and suggests that the evolution of legumin genes implicated loss of introns. From our data and from functional approaches recently described it becomes obvious that the posttranslational processing site of legumin precursors is less conserved than hitherto assumed. Finally, we present a phylogenetic analysis of legumin encoding sequences and discuss their utility as molecular markers for the reconstruction of seed plant evolution.
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  • 20
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    Journal of molecular evolution 41 (1995), S. 492-497 
    ISSN: 1432-1432
    Keywords: Primate ; Microsatellite ; Evolution ; Mutation process
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dimeric short tandem repeats are a source of highly polymorphic markers in the mammalian genome. Genetic variation at these hypervariable loci is extensively used for linkage analysis, for the identification of individuals, and may be useful for interpopulation and interspecies studies. In this paper, we analyze the variability and the sequences of a segment including three microsatellites, first described in man, in several species of primates (chimpanzee, orangutan, gibbon, and macaque) using the heterologous primers (man primers). This region is located on the human chromosome 6p, near the tumor necrosis factor genes, in the major histocompatibility complex. The fact that these primers work in all species studied indicates that they are conserved throughout the different lineages of the two superfamilies, the Hominoidea and the Cercopithecidea, represented by the macaques. However, the intervening sequence displays intraspecific and interspecific variability. The sites of base substitutions and the insertion/ deletion events are not evenly distributed within this region. The data suggest that it is necessary to have a minimal number of repeats to increase the rate of mutation sufficiently to allow the development of polymorphism. In some species, the microsatellites present single base variations which reduce the number of contiguous repeats, thus apparently slowing the rate of additional slippage events. Species with such variations or a low number of repeats are monomorphic. These microsatellite sequences are informative in the comparison of closely related species and reflect the phylogeny of the Old World monkeys, apes, and man.
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  • 21
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    Journal of molecular evolution 40 (1995), S. 405-412 
    ISSN: 1432-1432
    Keywords: Primate ; Evolution ; Heterochromatin ; Repetitive sequences ; β Satellite sequences ; L1 transposon ; Polymerase chain reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genomes of four primate species, belonging to the families Pongidae (chimpanzee, gorilla, and orangutan) and Hylobatidae (gibbons), have been analyzed for the presence and organization of two human GC-rich heterochromatic repetitive sequences: βSatellite (βSat) and LongSau (LSau) repeats. By Southern blot hybridization and PCR, both families of repeats were detected in all the analyzed species, thus indicating their origin in an ape ancestor. In the chimpanzee and gorilla, as in man, βSat sequences showed a 68-bp Sau3A periodicity and were preferentially organized in large clusters, whereas in the orangutan, they were organized in DNA fragments of 550 bp, which did not seem to be characterized by a tandem organization. On the contrary, in each of the analyzed species, the bulk of LSau sequences showed a longer Sau3A periodicity than that observed in man (450–550 bp). Furthermore, only in the chimpanzee genome some of LSau repeats seemed to be interspersed within blocks of βSat sequences. This sequence organization, which also characterizes the human genome, is probably absent in the gorilla. In fact, the analysis of a gorilla genomic library suggested that LSau repeats are not preferentially in linkage with βSat sequences. Moreover, LSau sequences were found in a genomic sector characterized by the simultaneous presence of L1 and (CA) repeats, as well as of anonymous sequences and known genes. In spite of the different sequence organization, the nucleotide differences between complete human and gorilla LSau repeats were very few, whereas one gorilla LSau repeat, interrupted by a probably-truncated L1 transposon, showed a higher degree of divergence. Besides the gorilla, this unusual sequence organization was detected in man, and, to a lesser extent, in the chimpanzee.
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  • 22
    ISSN: 1432-1432
    Keywords: Chloroplasts ; Evolution ; Rubisco ; Picoplankton ; Prochloron ; Prochlorothrix ; Prochlorococcus ; Prochlorales
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The prochlorophytes, oxygenic photosynthetic prokaryotes having no phycobiliprotein but possessing chlorophylls a and b, have been proposed to have a common ancestry with green chloroplasts, yet this is still controversal. We report here that partial sequence comparisons of the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase, including sequence data from two prochlorophytes, Prochlorococcus and Prochloron, indicate that Prochlorococcus is more closely related to a photosynthetic bacterium, Chromatium vinosum (γ-purple bacteria), than to cyanobacteria, while Prochloron is closely related to the prochlorophyte Prochlorothrix and to cyanobacteria. The molecular phylogenetic tree indicates that a common ancestor of Prochlorococcus and γ-purple bacteria branched off from the land plant lineage earlier than Prochloron, Prochlorothrix, and cyanobacteria.
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  • 23
    ISSN: 1432-1432
    Keywords: Alcohol dehydrogenase ; Caenorhabditis elegans ; Evolution ; Gene families
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have characterized two cDNA clones from the nematode Caenorhabditis elegans that display similarity to the alcohol dehydrogenase (ADH) gene family. The nucleotide sequences of these cDNAs predict that they encode Zn-containing long-chain ADH enzymes. Phylogenetic analysis suggests that one is most similar to dimeric class III ADHs found in diverse taxa; the other is most similar to the tetrameric forms of ADH previously described only in fungi.
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  • 24
    ISSN: 1432-1432
    Keywords: Cyanelles ; Cyanophora paradoxa ; Endosymbiosis ; Evolution ; Glaucocystophyta ; Glaucophyta ; Phylogeny ; Plastid ; 16S ribosomal RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Glaucocystophyte algae (sensu Kies, Berl. Deutsch. Bot. Ges. 92, 1979) contain plastids (cyanelles) that retain the peptidoglycan wall of the putative cyanobacterial endosymbiont; this and other ultrastructural characters (e.g., unstacked thylakoids, phycobilisomes) have suggested that cyanelles are “primitive” plastids that may represent undeveloped associations between heterotrophic “host” cells (i.e., glaucocystophytes) and cyanobacteria. To test the monophyly of glaucocystophyte cyanelles and to determine their evolutionary relationship to other plastids, complete 16S ribosomal RNA sequences were determined for Cyanophora paradoxa, Glaucocystis nostochinearum, Glaucosphaera vacuolata, and Gloeochaete wittrockiana. Plastid rRNAs were analyzed with the maximum-likelihood, maximumparsimony, and neighbor joining methods. The phylogenetic analyses show that the cyanelles of C. paradoxa, G. nostochinearum, and G. wittrockiana form a distinct evolutionary lineage; these cyanelles presumably share a monophyletic origin. The rDNA sequence of G. vacuolata was positioned within the nongreen plastid lineage. This result is consistent with analyses of nuclear-encoded rRNAs that identify G. vacuolata as a rhodophyte and support its removal from the Glaucocystophyta. Results of a global search with the maximumlikelihood method suggest that cyanelles are the first divergence among all plastids; this result is consistent with a single loss of the peptidoglycan wall in plastids after the divergence of the cyanelles. User-defined tree analyses with the maximum-likelihood method indicate, however, that the position of the cyanelles is not stable within the rRNA phylogenies. Both maximumparsimony and neighbor-joining analyses showed a close evolutionary relationship between cyanelles and nongreen plastids; these phylogenetic methods were sensitive to inclusion/exclusion of the G. wittrockiana cyanelle sequence. Base compositional bias within the G. wittrockiana 16S rRNA may explain this result. Taken together the phylogenetic analyses are interpreted as supporting a near-simultaneous radiation of cyanelles and green and nongreen plastids; these organelles are all rooted within the cyanobacteria.
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  • 25
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    Surveys in geophysics 16 (1995), S. 533-591 
    ISSN: 1573-0956
    Keywords: Satellites ; Giant planets ; Origin ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences , Physics
    Notes: Abstract The paper reviews the problem of formation and evolution of the so-called “regular satellites “ of the giant planets, and it consists of two parts: the first describes the possible origin of the satellites, the second studies their evolution, attempting to stress the relations of the present status of the satellites with their evolutionary history. The formation of regular satellite systems around giant planets is probably related to the formation of the central planet. Some characteristics of regular satellite systems are quite similar, and suggest a common origin in a disk present around the central body. This disk can originate through different mechanisms which we will describe, paying attention to the so-called “accretion disk” model, in which the satellite-forming material is captured. The disk phase links the formation of the primary body with the formation of satellites. The subsequent stages of the disk's evolution can lead first to the formation of intermediate size bodies, and through the collisional evolution of these bodies, to the birth of satellite “embryos” able to gravitationally capture smaller bodies. Given the scenario in which icy satellites may be formed by homogeneous accretion of planetesimals made of a mixtures of ice and silicates, if no melting occurs during accretion, the satellites have a homogeneous ice-rock composition. For the smaller satellites this homogeneous structure should not be substantially modified; only sporadic local events, such as large impacts, can modify the surface structure of the smaller satellites. For the larger satellites, if some degree of melting appears during accretion, a differentiation of the silicate part occurs, the amount of differentiation and hence the core size depending on the fraction of gravitational potential energy retained during the accumulation process. Melting and differentiation soon after the accretion, for the larger satellites, could also depend on the convective evolution in presence of phase transitions and generate an intermediate rock layer, considerably denser than the underlying, still homogeneous core, and unstable to overturning on a geologic time scale. Moreover the liquid water mantle could be a transient feature because the mantle would freeze over several hundred million years. For these large bodies the stable configuration is expected to be one consisting of a silicate core and a mantle of mixed rock and ice.
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  • 26
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    Development genes and evolution 205 (1995), S. 160-170 
    ISSN: 1432-041X
    Keywords: Drosophila ; Evolution ; fz ; Homeodomain ; Plasticity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Drosophila melanogaster segmentation gene fushi tarazu (ftz) encodes a homeodomain-type transcription factor involved in the control of larval pattern formation. Loss of function mutations cause an embryonic lethal, pair-rule phenotype. The segmentation defects, but not the lethality, can be partially rescued by the ftz orthologue from Drosophila hydei. In this work, the primary structure, expression and regulation of the D. hydei ftz gene was characterized. Sequence comparisons classify ftz as a rather fast evolving gene. However, since the homeodomain of the D. hydei FTZ protein is highly similar to that of D. melanogaster, proper regulation of D. melanogaster ftz downstream genes would be expected. In D. melanogaster embryos, a D. hydei ftz transgene is expressed normally, independent of endogenous ftz gene activity, suggesting that D. hydei ftz regulatory sequences are correctly recognized by D. melanogaster transcription factors. Accordingly, lacZ fusion constructs driven by the D. hydei ftz upstream element are expressed normally in D. melanogaster embryos. Altogether, the similarities between the two ftz orthologues by far outweigh the differences. The limited success of the trans-species rescue might be, therefore, a consequence of the accumulation of too many subtle changes in gene function, exceeding the limits of developmental plasticity during fly embryogenesis.
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  • 27
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    Sexual plant reproduction 8 (1995), S. 189-196 
    ISSN: 1432-2145
    Keywords: Endosperm balance number ; Congruity ; Imprinting ; Evolution ; Lycopersicon ; Triploid block
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Success of seed development following sexual crosses is primarily dependent on proper endosperm function and development. The failure to produce triploids, or “triploid block” in 4x×2x crosses served as the impetus for numerous studies of embryo and endosperm to attempt to explain cross failure. Early explanations were based upon a concept of a 2∶3∶2 ploidy balance between maternal tissue, endosperm, and embryo. Subsequent studies done with maize demonstrated that normal endosperm development in intraspecific maize crosses is dependent solely on having a 2∶1 maternal to paternal genome dosage in the endosperm. These results have been modified and extended to solanaceous species in the form of an endosperm dosage system in which empirically determined factors must bear the same 2∶1 relationship for crosses to succeed. Crossing behavior of these species suggest that the system is polygenically controlled and regulates both interspecific and intraspecific crosses. Endosperm dosage systems explain many aspects of species evolution, but the system appears to have originated as an ancient means of ensuring diploid fidelity.
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  • 28
    ISSN: 1432-0983
    Keywords: Green algae ; Volvox ; Transcription signals ; Gene expression ; Intron ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genome of the green alga Chlamydomonas reinhardtii contains approximately 15 gene clusters of the nucleosomal (or core) histone H2A, H2B, H3 and H4 genes and at least one histone H1 gene. Seven non-allelic histone gene loci were isolated from a genomic library, physically mapped, and the nucleotide sequences of three isotypes of each core histone gene species and one linked H1 gene determined. The core histone genes are organized in clusters of H2A–H2B and H3–H4 pairs, in which each gene pair shows outwardly divergent transcription from a short (300 bp) intercistronic region. These intercistronic regions contain typically conserved promoter elements, namely a TATA-box and the three motifs TGGCCAG-G(G/C)-CGAG, CGTTGACC and CGGTTG. Different from the genes of higher plants, but like those of animals and the related alga Volvox the 3′ untranslated regions contain no poly A signal, but a palindromic sequence (3′ palindrome) essential for mRNA processing is present. One single H1 gene was found in close linkage to a H2A–H2B pair. The H1 upstream region contains the octameric promoter element GGTTGA-CC (also found upstream of the core histone genes) and two specific sequence motifs that are shared only with the Volvox H1 promoters. This suggests differential transcription of the H1 and the core histone genes. The H1 gene is interrupted by two introns. Unlike Volvox H3 genes, the three sequenced H3 isoforms are intronfree. Primer-directed PCR of genomic DNA demonstrated, however, that at least 8 of the about 15 H3 genes do contain one intron at a conserved position. In synchronized C. reinhardtii cells, H4 mRNA levels (representative of all core histone mRNAs) peak during cell division, suggesting strict replication-dependent gene control. The derived peptide sequences place C. reinhardtii core histones closer to plants than to animals, except that the H2A histones are more animal-like. The peptide sequence of histone H1 is closely related to the V. carteri VH1-II (66% identity). Organization of the core histone gene in pairs, and non-polyadenylation of mRNAs are features shared with animals, whereas peptide sequences and enhancer elements are shared with higher plants, assigning the volvocalean histone genes a position intermediate between animals and plants.
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  • 29
    ISSN: 1432-0983
    Keywords: RNA editing ; Group-II intron ; Gene transfer ; Evolution ; tRNA genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A novel group II intron has been identified in the pea (Pisum sativum) mitochondrial genome. The gene harbouring this intron is identified as rps10 (encoding protein S10 of the small ribosomal subunit) by similarity to its known homologues in bacteria and in the mitochondrion of the liverwort Marchantia polymorpha. The rps10 gene is transcribed in pea, the intron is removed, and RNA editing in the rps10 reading frame increases similarity to its homologue in the M. polymorpha mitochondrion. Contrary to the situation in bacteria and Marchantia, rps10 is not part of a ribosomal-protein gene cluster in pea. It is flanked upstream by the genes trnF and trnP, encoding phenylalanine-and proline-accepting tRNAs, and downstream by cox1, encoding subunit 1 of the cytochrome-c-oxidase. Southern hybridization shows that sequences homologous to rps10 exist in potato mitochondria but not in mitochondria of Oenothera berteriana and Arabidopsis thaliana. The pea rps10 intron is homologous to introns in rrn26 and cox3 in the Marchantia mitochondrial genome, while the Marchantia rps10 gene lacks an intron.
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  • 30
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    Current genetics 28 (1995), S. 410-414 
    ISSN: 1432-0983
    Keywords: Mitochondrial DNA ; Multi-copy ; Pseudogene ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four nuclear pseudogenes homologous to the 10031–10195-bp region of the human mitochondrial genome were detected by constant denaturant capillary electrophoresis. Among them, one pseudogene is present as at least five copies in each cell, in accordance with our previous observations of multi-copy mitochondrial DNA pseudogenes. The presence of multiple identical copies of pseudogenes suggests that the human genome underwent a series of genetic changes, including gene amplifications, very recently in evolutionary history, i.e., within the last 390000 years.
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    Current genetics 28 (1995), S. 97-99 
    ISSN: 1432-0983
    Keywords: GIY-YIG ORF ; Endonuclease ; Plastid ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Group-I introns, containing open reading frames (ORFs) that code for homing endonucleases, are widely distributed amongst eukaryotic organellar genomes. However, endonucleases of the GIY-YIG subclass have a restricted distribution in mitochondria and bacteriophages, and have never been observed in plastids. We have found the GIY-YIG motif in an intronic ORF within the previously published psbA gene sequence from Chlamydomonas reinhardtii chloroplasts. Based on phylogenetic analysis and an evaluation of amino-acid substitutions, this ORF is not closely related to any of the other GIY-YIG ORFs. These results suggest that GIY-YIG ORFs have a longer evolutionary history than previously assumed.
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  • 32
    ISSN: 1432-072X
    Keywords: Magnetospirillum magnetotacticum ; Cytochrome c ; Amino acid sequence ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cytochrome c-550 was purified from Magnetospirillum magnetotacticum to an electrophoretically homogeneous state, and some of its properties were determined. The cytochrome showed absorption peaks at 528 and 409 nm in the oxidized form, and at 550, 521, and 414 nm in the reduced form. Its midpoint redox potential at pH 7.0 was determined to be +289 mV. The primary structure of cytochrome c-550 was determined. Cytochrome c is composed of 97 amino acid residues, and its molecular weight was calculated to be 10,873, including heme c. Its primary structure is very similar to those of Rhodospirillum fulvum and Rhodospirillum molischianum cytochromes c 2, suggesting that M. magnetotacticum is phylogenetically related to photosynthetic bacteria.
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  • 33
    ISSN: 1432-072X
    Keywords: Key wordsMagnetospirillum magnetotacticum ; Cytochrome c ; Amino acid sequence ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cytochrome c-550 was purified from Magnetospirillum magnetotacticum to an electrophoretically homogeneous state, and some of its properties were determined. The cytochrome showed absorption peaks at 528 and 409 nm in the oxidized form, and at 550, 521, and 414 nm in the reduced form. Its midpoint redox potential at pH 7.0 was determined to be +289 mV. The primary structure of cytochrome c-550 was determined. Cytochrome c is composed of 97 amino acid residues, and its molecular weight was calculated to be 10,873, including heme c. Its primary structure is very similar to those of Rhodospirillum fulvum and Rhodospirillum molischianum cytochromes c 2, suggesting that M. magnetotacticum is phylogenetically related to photosynthetic bacteria.
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  • 34
    ISSN: 1573-4919
    Keywords: vasoactive intestinal peptide ; ulcerative colitis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The availability of colon provides a ready source of human neurons. Among the products of nerve cell bodies, vasoactive intestinal peptide is a neuropeptide that serves as a marker of non-adrenergic, non-cholinergic inhibitory nerves in colon. These nerves have been proposed to be involved in regulation of immune function, secretion, and smooth muscle function. In previous work, we identified decreased tissue levels of vasoactive intestinal peptide in a disorder of chronic colonic mucosal inflammation, ulcerative colitis. We hypothesized that diminished gene expression of vasoactive intestinal peptide could result in decreased tissue levels of this neuropeptide. Sigmoid colon was obtained at surgery from controls (n=6) and patients with ulcerative colitis (n=6). Vasoactive intestinal peptide mRNA was quantified by Northern blot hybridization and tissue levels of vasoactive intestinal peptide were determined by radioimmunoassay. Tissue vasoactive intestinal peptide was decreased only in the mucosalsubmucosal layer of ulcerative colitis (p=.02). There was a single 1.7 kbase vasoactive intestinal peptide transcript identified in both control colon and ulcerative colitis. Normalized vasoactive intestinal peptide mRNA levels were increased by 260% in ulcerative colitis compared to controls (p〈.01). These observations suggest that decreased vasoactive intestinal peptide gene expression or abnormal post-transcriptional processing are not primary defects in this disorder of chronic inflammation. The findings support the alternative hypothesis that axonal degeneration in ulcerative colitis could result in increased expression of neuronal vasoactive intestinal peptide mRNA.
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  • 35
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; insulin ; calcium ; gene expression ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of refeeding on the expression of Ca2+-binding protein regucalcin mRNA in the liver of fasted rats was investigated. When rats were fasted overnight, the hepatic regucalcin mRNA level was reduced about 70% of that in feeding rats. Refeeding produced a remarkable elevation of hepatic regucalcin mRNA level (about 150–170% of fasted rats). Liver regucalcin concentration was appreciably increased by refeeding, although it was not altered by fasting. The oral administration of glucose (2 g/kg body weight) to fasted rats caused a significant increase in hepatic regucalcin mRNA level. Moreover, hepatic regucalcin mRNA level was clearly elevated by a single subcutaneous administration of insulin (10 and 100 U/kg) to fasted rats. The hormonal effect was not further enhanced by the simultaneous administration of calcium chloride (250 mg Ca/kg) to fasted rats, although calcium administration stimulated regucalcin mRNA expression in the liver. The present study suggests that the expression of hepatic regucalcin mRNA stimulated by refeeding is significantly involved in the action of insulin and/or calcium as stimulating factors.
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    Molecular and cellular biochemistry 143 (1995), S. 67-71 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; gene distribution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The existence and expression of gene encoding the Ca2+-binding protein regucalcin in various species and tissues were investigated with Southern and Northern hybridization analyses using regucalcin cDNA (0.9 kb of open reading frame). Genomic Southern hybridization analysis demonstrated that regucalcin gene was widely conserved among higher animals including human, monkey, rat, mouse, dog, bovine, rabbit and chicken. The gene was not found in yeast. The Northern blot analysis of poly (A)+RNAs extracted from the liver of various species showed that regucalcin mRNA was predominantly expressed in rat and mouse, although the expression was also seen in human, bovine and chicken. Furthermore, the enzyme-linked immunoadsorbent assay (ELISA) with rabbit-anti-regucalcin IgG indicated that hepatic regucalcin concentration was most pronounced in rat as compared with that of guinea pig, mouse and chicken. These observations show that the gene expression of regucalcin and its protein synthesis is unique in the liver of rats, suggesting the existence of a specific mechanism in demonstrating regucalcin synthesis from gene.
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    Molecular and cellular biochemistry 143 (1995), S. 137-141 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; estrogen ; gene expression ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of nuclear receptor-related hormones on the expression of hepatic calcium-binding protein regucalcin mRNA in rats was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.9 kb of open-reading frame). A single subcutaneons administration of 17β-estradiol (0.5, 1.0 and 2.0 mg/kg body weight) in rats induced a remarkable increase of regucalcin mRNA in liver; the level was about 200% of control at 24 h after the administration of 2.0 mg/kg. The increase showed about 350% even at 6 h after the administration. Meanwhile, hepatic regucalcin mRNA level was not appreciably altered by a single subcutaneous administration of thyroxine (T4) (20, 40 and 80 mg/kg) or hydrocortisone (10 and 30 mg/kg) in rats. The present study demonstrates that the expression of hepatic regucalcin mRNA is stimulated by estrogen action in the liver nuclei of rats.
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    Molecular and cellular biochemistry 144 (1995), S. 105-108 
    ISSN: 1573-4919
    Keywords: fatty acid synthase ; gene expression ; and thyroid hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of triiodothyronine (T3) on regulation of fatty acid synthase in chicken liver was investigated. In hypothyroid animals, enzyme activity was about one half of that in euthyroid animals. T3 treatment increased the enzyme activity in hypothyroid animals. There is little difference in both the mRNA concentration and the transcription rate between euthyroid and hypothyroid animals. T3 treatment markedly decreased both the mRNA concentration and the transcription rate in euthyroid and hypothyroid animals. These results suggested that T3 maintained the normal level of enzyme expression primarily by stimulating the post-transcriptional step, while the transcription of the gene was inhibited by hyperthyroidism.
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    Molecular and cellular biochemistry 148 (1995), S. 45-57 
    ISSN: 1573-4919
    Keywords: manganese ; superoxide dismutase ; gene expression ; hyperoxide lung injury ; nuclear factor kappa B
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract TNFα and IL-1 each can activate NF-κB and induce gene expression of manganese superoxide dismutase (MnSOD), a mitochondrial matrix enzyme which can provide critical protection against hyperoxic lung injury. The regulation of MnSOD gene expression is not well understood. Since redox status can modulate NF-κB and potential κB site(s) exist in the MnSOD promoter, the effect of thiols (including NAC, DTT and 2-ME) on TNFα and IL-1 induced activation of NF-κB and MnSOD gene expression was investigated. Activation of NF-kB and increased MnSOD expression were potentiated by thiol reducing agents. In contrast, thiol oxidizing or alkylating agents inhibited both NF-κB activation and elevated MnSOD expression in response to TNFα or IL-1. Since protease inhibitors TPCK and TLCK can inhibit NF-κB activation, we also investigated the effect of these compounds on MnSOD expression and NF-κB activation. TPCK and TLCK each inhibited MnSOD gene expression and NF-κB activation. Since the MnSOD promoter also contains anAP-1 binding site, the effect of thiols and thiol modifying agents on AP-1 activation was investigated. Thiols had no consistent effect onAP-1 activation. Likewise, some of the thiol modifying compounds inhibited AP-1 activation by TNFα or IL-1, whereas others did not. Since diverse agents had similar effects on activation of NF-κB and MnSOD gene expression, we have demonstrated that activation of NF-κB and MnSOD gene expression are closely associated and that reduced sulfhydryl groups are required for cytokine mediation of both processes.
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    Molecular and cellular biochemistry 146 (1995), S. 71-77 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; rat kidney cortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb of open-reading frame). Regucalcin mRNA was expressed in the kidney cortex, and this expression was clearly increased by a single intraperitoneal administration of calcium chloride solution (5–15 mg Ca/100 g body weight) in rats; this increase was remarkable at 60–120 min after the administration. Thyroparathyroidectomy (TPTX) caused a slight decrease of regucalcin mRNA levels in the kidney cortex. However, the administration of calcium (10 mg/100 g) in TPTX rats produced a clear increase of regucalcin mRNA levels in the kidney cortex. The subcutaneous administration of calcitonin (10–100 MRC mU/100 g) or parathyroid hormone [1–34] (1–10 U/100 g) in TPTX rats which received calcium (10 mg/100 g) administration did not cause an appreciable alteration of regucalcin mRNA levels in the kidney cortex, suggesting that the mRNA expression is not stimulated by calcium-regulating hormones. The administration of trifluoperazine (TFP; 5 mg/100 g), an inhibitor of Ca2+/calmodulin action, completely blocked the expression of regucalcin mRNA stimulated by calcium administration. Now, calcium content in the kidney cortex was significantly elevated by a single intraperitpneal administration of calcium (10 mg/100 g) in rats. The present study clearly demonstrates that the expression of regucalcin mRNA in the kidney cortex is stimulated by calcium administration in rats. This expression may be mediated through Ca2+/calmodulin action in the kidney cortex.
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    Astrophysics and space science 230 (1995), S. 169-176 
    ISSN: 1572-946X
    Keywords: Late-type Stars ; Evolution ; Carbon Stars ; RV Tauri Stars
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract The M, S and C stars may be placed in an evolutionary sequence on the basis of direct observation of the spectroscopic transitions on the AGB of rich intermediate-age clusters in the Magellanic Clouds, but some S and C stars cannot be accounted for in this way. The S stars inω Centauri owe their peculiarity to a primordial enrichment in s-process elements. The J-type (13C-rich) carbon stars originate in a different way to the ordinary cool N-type carbon stars. Some of them have silicate-rich circumstellar dust, contrary to expectation. Some of the carbon-rich RV Tauri stars also have silicate-rich dust and in both cases it may be organised in a disc. Observational evidence for gas and dust ejection by ordinary N-type carbon stars has been found and may be inferred for some RV Tauri stars.
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    Astrophysics and space science 224 (1995), S. 29-42 
    ISSN: 1572-946X
    Keywords: Protostars ; Evolution ; Infall ; Jets
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract The use of sensitive receivers on large ground-based radiotelescopes such as the JCMT, the IRAM 30 m MRT, and the VLA has recently yielded significant progress in our observational understanding of low-mass protostars. Submillimeter continuum observations suggest that the youngest stellar objects detected in the near-/mid-IR range -the so-called Class I sources or “infrared protostars” - have only residual amounts of circumstellar material and are thus relatively evolved. At the same time, a smaller number of colder and more obscured YSOs - designated “Class 0” - characterized by virtually no emission below 10µm but strong submillimeter emission have been identified. These Class 0 or “submillimeter protostars” have not yet assembled the bulk of their final stellar mass, and correspond to the youngest protostar stage known to date (probable age ≲ 104 yr). Direct evidence for gravitational infall has been found in some of these sources confirming their protostellar nature. However, most (if not all) Class 0 protostars already drive highly collimated CO outflows.
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    Molecular and cellular biochemistry 151 (1995), S. 55-60 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium ; gene expression ; kidney damage ; rat kidney cortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The alteration of Ca2+-binding protein regucalcin mRNA expression in the kidney cortex of rats administered cisplatin and cephaloridine, which can induce kidney damage, was investigated. Cisplatin (0.25, 0.5 and 1.0 mg/100 g body weight) or cephaloridine (25, 50 and 100 mg/100 g) was intraperitoneally administered in rats, and 1, 2 and 3 days later they were sacrificed. The alteration in serum findings after the administration of cisplatin (1.0 mg/100 g) or cephaloridine (50 and 100 mg/100 g) demonstrated chemically induced kidney damage; blood urea nitrogen (BUN) concentration increased markedly and serum inorganic phosphorus or calcium concentration decreased significantly. Moreover, the administration of cisplatin (1.0 mg/100 g) or cephaloridine (100 mg/100 g) caused a remarkable increase of calcium content in the kidney cortex of rats, indicating kidney damage. The expression of regucalcin mRNA in the kidney cortex was markedly reduced by the administration of cisplatin or cephaloridine in rats, when the mRNA levels were analyzed by Northern blotting using rat liver regucalcin cDNA (0.9 kb). The mRNA decreases were seen with the used lowest dose of cisplatin or cephaloridine. The present study clearly demonstrates that the mRNA expression of Ca2+-binding protein regucalcin in the kidney cortex of rats is decreased by chemically induced kidney damage.
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  • 44
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    Molecular and cellular biochemistry 152 (1995), S. 131-141 
    ISSN: 1573-4919
    Keywords: gene expression ; mRNA ; proto-oncogenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Polyomavirus large T-antigen transgenic mice develop cardiac hypertrophy characterized by an increase in atrial natriuretic factor and β-myosin heavy chain isoform expression. The aim of this study was to examine changes in proto-oncogene expression in hypertrophied hearts from the transgenic mice. Expression of early growth response-1 (Egr-1) mRNA was detected in hearts from all 15 transgenic mice, but was not detectable in 13 control mice. Reverse transcriptase-polymerase chain reaction experiments usingEgr-1-specific primers confirmed the increase inEgr-1 mRNA in enlarged hearts from the transgenic mice. Expression of c-jun,junD and Ha-ras mRNAs was increased in the transgenic hearts 3, 17 and 2.8-fold, respectively. Western blots showed an increase in c-myc, c-jun and ras protein in hypertrophied transgenic hearts. Immunofluorescence analyses confirmed an increase in Egr-1 and c-jun protein in transgenic cardiomyocytes. Proliferating cell nuclear antigen, Ki-ras and HSP 90 mRNAs were decreased 22, 2.7 and 3-fold, respectively in the transgenic hearts. Not altered in most hypertrophied hearts was expression of c-fos, junB, p53, c-neu, c-myc, HSP70, HSP27, TGF-β or IGF-1 mRNAs. Proto-oncogene and growth factor gene expression in hypertrophy induced by PVLT expression is modulated, with some proto-oncogenes increased and others decreased in expression.
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  • 45
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    Journal of computer aided molecular design 9 (1995), S. 181-202 
    ISSN: 1573-4951
    Keywords: De novo drug design ; Evolution ; Automated structure generation ; 3D database ; Expert system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A genetic algorithm has been designed which generates molecular structures within constraints. The constraints may be any useful function, for example an enzyme active site, a pharmacophore or molecular properties from pattern recognition or rule-induction analyses. The starting point may be random or may utilise known molecules. These are modified to ‘grow’ into families of structures which, using the evolutionary operators of selection, crossover and mutation evolve to better fit the constraints. The basis of the algorithm is described together with some applications in lead generation, 3D database construction and drug design. Genetic algorithms of this type may have wider applications in chemistry, for example in the design and optimisation of new polymers, materials (e.g. superconducting materials) or synthetic enzymes.
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  • 46
    ISSN: 1573-5028
    Keywords: β-fructofuranosidase ; invertase ; gene expression ; gene structure ; flower buds ; Daucus carota
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three genomic clones (Inv *Dc1, Inv *Dc2 and Inv *Dc3) were isolated by using the cDNA for carrot cell wall β-fructofuranosidase as a probe. The expression patterns of the three genes differed markedly. High levels of Inv *Dc1 transcripts were found in leaves and roots of young carrot, whereas in plants with developing tap roots no transcripts were detected. A high level of mRNA of Inv *Dc1 was also present in suspension-cultured cells. In developing reproductive organs, only low levels of transcripts of Inv *Dc1 were found in flower buds and flowers and none at later stages of development. In contrast, Inv *Dc2 and Inv *Dc3 were not expressed in vegetative plant organs. Invb1 *Dc1 was exclusively and strongly expressed in flower buds, and Inv *Dc3 at a very low level in suspension-cultured cells.
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  • 47
    ISSN: 1573-5028
    Keywords: fruit ; gene expression ; promoter ; ripening ; tomato ; transgenic plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 1.4 kb 5′ polygalacturonase (PG) gene-flanking region has previously been demonstrated to direct ripening-specific chloramphenicol acetyl transferase (CAT) expression in transgenic tomato plants. The steady state level of CAT mRNA in these plants was estimated to be less than 1% of the endogenous PG mRNA. Further constructs containing larger PG gene-flanking regions were generated and tested for their ability to direct higher levels of reporter gene expression. A 4.8 kb 5′-flanking region greatly increased levels of ripening-specific reporter gene activity, while a 1.8 kb 3′ region was only shown to have a positive regulatory role in the presence of the extended 5′ region. Transgenic plants containing the CAT gene flanked by both of these regions showed the same temporal pattern of accumulation of CAT and PG mRNA, and steady-state levels of the transgene mRNA were equivalent to 60% of the endogenous PG mRNA on a per gene basis. The proximal 150 bp of the PG promoter gave no detectable CAT activity. However, the distal 3.4 kb of the 4.8 kb 5′ PG promoter was shown to confer high levels of ripening-specific gene expression when placed in either orientation upstream of the 150 bp minimal promoter. The DNA sequence of the 3.4 kb region revealed a 400 bp imperfect reverse repeat, and sequences which showed similarity to functionally significant sequences from the ripening-related, ethylene-regulated tomato E8 and E4 gene promoters. The possible roles of the flanking regions in regulating PG gene expression are discussed.
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  • 48
    ISSN: 1573-5028
    Keywords: acetohydroxyacid synthase ; gene organization ; gene expression ; herbicide resistance ; cotton ; Gossypium hirsutum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The acetohydroxyacid synthase (AHAS) gene family of the cotton AD allotetraploid Gossypium hirsutum has been cloned and characterized. We have identified six different AHAS genes from an analysis of genomic clones and Southern blots of genomic DNA. Four of the six genes are organized as tandem pairs, in which the genes are separated by only 2–3 kb. Conservation of restriction fragment length polymorphisms between G. hirsutum and A-genome and D-genome-containing diploid cottons was sufficient to assign the single genes in clones A5 and A19 to the A and D subgenomes, respectively. Each diploid genome has one tandem pair, but in these cases we could not make specific subgenomic assignments. DNA and deduced amino acid sequences were determined for the A5 and A19 genes, and an AHAS cDNA clone isolated from a leaflibrary. The sequence of the A19 gene matches that of the cDNA clone, while the A5 gene is 97.8% similar. The four genes comprising the tandem pairs are much less similar to the cDNA clone. The deduced amino acid sequences of the mature polypeptides encoded by the A5 and A19 genes are collinear with the housekeeping forms of AHAS from Arabidopsis thaliana, Nicotiana tabacum and Brassica napus. The constitutive expression of A5 and A19 was confirmed with RNase protection assays and northern blots. We conclude that these genes encode the main house-keeping froms of AHAS in G. hirsutum. Among the four AHAS genes comprising the two tandem pairs, at least two are functional. These genes exhibit either low-level constitutive expression (one or both of the ‘downstream’ genes of each pair), or highly specific expression in reproductive tissue (one or both of the ‘upstream’ genes of each pair). The AHAS gene family of G. hirsutum is more complex than that of other plants so far examined.
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  • 49
    ISSN: 1573-5028
    Keywords: Brassica ; phosphate starvation ; gene expression ; β-glucosidase ; mineral nutrition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Differential gene expression at the transcriptional level was examined as an initial step in the investigation of the Pi starvation response of Brassica nigra suspension cells. Total RNA was extracted from 7-day old cells grown in media containing either no Pi, 1.25 mM or 10 mM Pi., In vitro translation was carried out using their respective poly(A)+ RNA isolates and the resultant polypeptides were separated on a high-resolution SDS-PAGE gel. Scanning densitometry identified four polypeptides (ca. 31.7, 32.3, 52.5 and 64.8 kDa) present only in the Pi-starved samples. Screening by differential hybridization was performed on a cDNA library constructed from mRNA isolated from Pi-starved cells. Probes prepared from mRNA from Pi-deficient and Pi-sufficient cells identified a number of clones representing mRNA species that were preferentially transcribed under Pi deficiency. These phosphate starvation-responsive (psr) clones were placed into eleven groups as determined by cross-hybridization. Northern blots showed that the corresponding genes are inducible in both mild and severe Pi starvation conditions. Preliminary sequencing identified one of the clones as being homologous to β-glucosidases from several plant species. The possible role of β-glucosidase during Pi starvation and the identities of the other psr genes are discussed.
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  • 50
    ISSN: 1573-5028
    Keywords: linked gene ; gene expression ; peroxidase ; Populus kitakamiensis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genomic library was prepared from Populus kitakamiensis and screened with the cDNA for an anionic peroxidase from P. kitakamiensis. One genomic clone was isolated that contained two tandemly oriented genes for anionic peroxidases, prxA3a and prxA4a. Both genes consisted of four exons and three introns; the introns had consensus nucleotides, namely, GT and AG, at their 5′ and 3′ ends, respectively. The prxA3a and prxA4a genes encoded 347 and 343 amino acid residues, respectively, including putative signal sequences at the amino-termini. Putative promoters and polyadenylation signals were found in the flanking regions of both genes. The sequence of the coding region of prxA3a was completely identical to that of the cDNA clone pA3, whereas the sequence of the coding region of prxA4a was only 73% identical to that of the cDNA clone pA3. Northern blot analysis showed that the patterns of expression of the mRNAs that corresponded to prxA3a and prxA4a differed in stems of P. kitakamiensis.
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  • 51
    ISSN: 1573-5028
    Keywords: anthocyanin ; Compositae ; corolla ; dfr ; flower development ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the ornamental cut flower plant Gerbera hybrida the spatial distribution of regulatory molecules characteristic of differentiation of the composite inflorescence is visualized as the various patterns of anthocyanin pigmentation of different varieties. In order to identify genes that the plant can regulate according to these anatomical patterns, we have analysed gene expression affecting two enzymatic steps, chalcone synthase (CHS) and dihydroflavonol-4-reductase (DFR), in five gerbera varieties with spatially restricted anthocyanin pigmentation patterns. The dfr expression profiles vary at the levels of floral organ, flower type and region within corolla during inflorescence development according to the anthocyanin pigmentation of the cultivars. In contrast, chs expression, although regulated in a tissue-specific manner during inflorescence development, varies only occasionally. The variation in the dfr expression profiles between the varieties reveals spatially specific gene regulation that senses the differentiation events characteristic of the composite inflorescence.
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  • 52
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    Plant molecular biology 27 (1995), S. 441-456 
    ISSN: 1573-5028
    Keywords: HSP70 ; HSC70 ; seed development ; imbibition ; chaperone ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Eukaryotes express several cytoplasmic HSP70 genes, and their encoded proteins participate in diverse cellular processes. Three cDNAs encoding highly expressed cytoplasmic HSP70 homologues from Pisum sativum were cloned and characterized. They were designated PsHSP71.2, PsHSC71.0, and PsHSP70b. These HSP70 genes have different expression profiles in leaves: PsHSP71.2 is observed only in response to heat stress, PsHSC71.0 is present constitutively, and PsHSP70b is weakly constitutively expressed, but induced strongly in response to heat stress. In addition to being heat induced, the PsHSP71.2 mRNA is also expressed in zygotic, but not maternal organs of developing pea seeds, while PsHSC71.0 and PsHSP70b mRNAs are present in maternal and zygotic organs throughout seed development. Immunoblot analysis of parallel protein samples detects a 70 kDa polypeptide in all samples, and a 72 kDa polypeptide that corresponds to the PsHSP71.2 gene product is observed in cotyledons beginning at mid-maturation and in axes beginning between late maturation and desiccation. This polypeptide is not detected in the seed coat. The 72 kDa polypeptide remains abundant in both cotyledons and axes through germination, but declines substantially between 48 and 72 h after the onset of imbibition. Differential control of HSP70 expression during heat stress, seed maturation, and germination is consistent with the hypothesis that there are functional distinctions between cytoplasmic HSP70s.
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  • 53
    ISSN: 1573-5028
    Keywords: Cucumis sativus ; gene expression ; glyoxylate cycle ; glyoxysome ; isocitrate lyase ; seed germination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cucumber (Cucumis sativus L.) genome contains only a single gene encoding the glyoxylate cycle enzyme isocitrate lyase (ICL). The cucumber icl gene has been isolated and sequenced, revealing only two small introns. The predicted amino acid sequence is more than 85% identical to ICL from other higher plants, and contains the C-terminal tripeptide Ser-Arg-Met which resembles a peroxisomal targeting sequence. The icl gene is coordinately expressed with the malate synthase (ms) gene after seed germination in both the light and the dark, suggesting that these genes may contain similar DNA elements regulating transcription. The start of transcription of the icl gene was determined and the DNA sequences upstream compared with the region of the ms gene promoter known to regulate transcription. This comparison revealed a highly conserved DNA sequence at similar positions in each gene.
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  • 54
    ISSN: 1573-5028
    Keywords: Alfalfa ; cell division cycle ; chromosomal location ; cyclin ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyclins in association with the protein kinase p34cdc2and related cyclin-dependent protein kinases (cdks) are key regulatory elements in controlling the cell division cycle. Here, we describe the identification and characterization of a full-length cDNA clone of alfalfa mitotic cyclin, termed CycIIIMs. Computer analysis of known plant cyclin gene sequences revealed that this cyclin belongs to the same structural group as the other known partial alfalfa cyclin sequences. Genetic segregation analysis based on DNA-DNA hybridization data showed that the CycIIIMs gene(s) locates in a single chromosomal region on linkage group 5 of the alfalfa genetic map between RFLP markers UO89A and CG13. The assignment of this cyclin to the mitotic cyclin class was based on its cDNA-derived sequence and its differential expression during G2/M cell cycle phase transition of a partially synchronized alfalfa cell culture. Sequence analysis indicated common motifs with both the A- and B-types of mitotic cyclins similarly to the newly described B3-type of animal cyclins.
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  • 55
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    Plant molecular biology 29 (1995), S. 647-662 
    ISSN: 1573-5028
    Keywords: gene expression ; peroxidase ; powdery mildew ; splicing ; Triticum aestivum L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A PCR-based screening approach was used to isolate genomic clones from wheat encoding peroxidase isozymes. Three complete genes (pox1, pox2 and pox4) and one truncated gene (pox3) were characterized. The nucleotide sequences predicted mature proteins of 31 kDa, in which all the highly conserved motifs of secreted plant peroxidases were preserved. The coding regions showed 73–83% DNA sequence identity, with the highest level of similarity noted for the tandemly oriented pox2 and pox3. Expression of respective pox genes in various tissues of wheat was assessed by the RT-PCR technique, which showed that all four genes are active. The primary pox1 mRNA was spliced to remove three introns, whereas processing of the other pox transcripts involved only two intervening sequences. Splicing occurred at consensus GU/AG splice sites except for the first introns of pox1, pox2 and pox4 transcripts, where processing took place at unusual GC donor sites. The RNA analysis suggested that the pox1, pox2 and pox4 genes are predominantly expressed in roots. Lower levels of expression were found for pox4 and pox3 in leaves. Infection of wheat by the powdery mildew fungus selectively induced expression of pox2 in leaves.
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  • 56
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    Plant molecular biology 29 (1995), S. 1211-1221 
    ISSN: 1573-5028
    Keywords: ferredoxin ; Citrus ; ethylene ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The sequence and expression of mRNA homologous to a cDNA encoding a non-photosynthetic ferredoxin (Fd1) from Citrus fruit was investigated. The non-photosynthetic nature of this ferredoxin was deduced from: (1) amino acid sequence alignments showing better scores with non-photosynthetic than with photosynthetic ferredoxins, (2) higher expression in tissues containing plastids other than chloroplast such as petals, young fruits, roots and peel of fully coloured fruits, and (3) the absence of light-dark regulation characteristic of photosynthetic ferredoxins. In a phylogenetic tree constructed with higher-plant ferredoxins, Citrus fruit ferredoxin clustered together with root ferredoxins and separated from the photosynthetic ferredoxins. Non photosynthetic (root and fruit) ferredoxins, but not the photosynthetic ferredoxins, have their closest homologs in cyanobacteria. Analysis of ferredoxin genomic organization suggested that non-photosynthetic ferredoxins exist in Citrus as a small gene family. Expression of Fd1 is developmentally regulated during flower opening and fruit maturation, both processes may be mediated by ethylene in Citrus. Exogenous ethylene application also induced the expression of Fd1 both in flavedo and leaves. The induction of non-photosynthetic ferredoxins could be related with the demand for reducing power in non-green, but biosynthetically active, tissues.
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  • 57
    ISSN: 1573-4978
    Keywords: MAR ; SCS ; insulation ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The flanking sequences of several genes have been shown to direct a position independent expression of transgenes. Attempts to completely identify the insulating sequences have failed so far. Some of these sequences contain a matrix attached region (MAR) located in the flanking part of the genes. This article will show that the MARs in cultured cells located in the 3' OH region of the human apolipoprotein B100 (Apo B100) and within the SV40 genome were unable to stimulate and insulate transgene expression directed by the promoters from a rabbit whey acidic protein (WAP) gene or from human cytomegalovirus (hCMV) early genes. In transgenic mice, the MAR from the Apo B100 and SV40 genes did not enhance the expression of a transgene containing the rabbit whey acid protein (WAP) promotor, the late gene SV40 intron (VP1 intron), the bovine growth hormone (bGH) cDNA and the SV40 late gene terminator. This construct was even toxic for embryos. Similarly, the specialized chromatin structure (SCS) from the Drosophila 87A7 HSP70 gene reduced chloramphenicol acetyl transferase (CAT) activity when added between a cytomegalovirus (CMV) enhancer and a Herpes simplex thymidine kinase (TK) gene promoter. This inhibitory action was almost complete when a second SCS sequence was added before the CMV enhancer. Sequences from the firefly luciferase and from the human gene cathepsin D cDNA used as control unexpectedly showed a similar inhibitory effect when added to the CMVTKCAT construct instead of SCS. When added before the CMV enhancer and after the transcription terminator in the CMVTKCAT construct, the SCS sequence was unable to insulate the integrated gene as seen by the fact that the level of CAT in cell extracts were by no means correlated with the number of copies in individual clones. From these data, it is concluded that i) a MAR containing the canonical AT rich sequences does not amplify the expression of all gene constructs ii) AT rich MAR sequences do not have per se an insulating effect iii) Drosophila SCS from the 87A7 HSP70 gene has no insulating effect in all gene constructs (at least in mammalian cells) iv) and the addition of a DNA fragment between an enhancer and a promoter in a gene construct cannot be used as a reliable test to evaluate its insulating property.
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  • 58
    ISSN: 1573-5028
    Keywords: gene expression ; embryo sac ; ovule ; Petunia hybrida ; protein ; kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mitogen activated protein (MAP) kinase pathway of eukaryotes is stimulated by many growth factors and is required for the integration of multiple cellular signals. In order to study the function of MAP kinases during plant ovule development we have synthesized a Petunia hybrida ovule-specific cDNA library and screened for MAP protein kinase-related sequences using a DNA probe obtained by PCR. A full-length cDNA clone was identified (PMEK for Petunia hybrida MAP/ERK-related protein kinase) and shown to encode a protein related to the family of MAP/ERK protein kinases. Southern blot analysis showed that PMEK is a member of a small multigene family in P. hybrida. The cDNA codes for a protein (PMEK1) of 44.4 kDa with an overall sequence identity of 44% to the products of the mammalian ERK/MAP kinase gene, and the budding yeast KSS1 and FUS3 genes. PMEK1 displays 96 and 80% identity respectively with the tobacco NTF3 and Arabidopsis ATMPK1 kinases, and only 50% to the more distantly related plant MAP kinase MsERK1 from alfalfa. The two phosphorylation sites found in the loop between subdomain VII and VIII in all the other MAP kinases are also present in PMEK1. RNA gel blot and RT-PCR analyses demonstrated that PMEK1 is expressed in vegetative organs and preferentially accumulated in female reproductive organs of P. hybrida. In situ hybridization experiments showed that in the reproductive organs PMEK1 is expressed only in the ovary and not in the stamen.
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  • 59
    ISSN: 1573-5028
    Keywords: ADP-glucose pyrophosphorylase (small and large subunit) ; DNA sequence ; gene expression ; starch synthesis ; sugar beet (Beta vulgaris L.)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cDNA cloning of a small and a large subunit of ADP-glucose pyrophosphorylase (AGPase) from sugar beet is reported. The deduced amino acid sequences are highly homologous to previously identified AGPase polypeptides from other plant species. Both subunits are encoded by low copy genes. When RNA gel blot experiments were performed, strongest expression was detected in sink and source leaves of greenhouse-grown sugar beet plants. A lower expression was found in other tissues tested, i.e. in the hypocotyl, the tap root and roots. In these tissues, slightly higher transcript levels were found for the small subunit gene than for the large subunit gene.
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  • 60
    ISSN: 1573-5028
    Keywords: glycine betaine ; betaine aldehyde dehydrogenase ; osmotic stress ; gene expression ; plant hormone ; abscisic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When subjected to salt stress or drought, some vascular plants such as barley respond with an increased accumulation of the osmoprotectant glycine betaine (betaine), being the last step of betaine synthesis catalyzed by betaine aldehyde dehydrogenase (BADH). We report here cloning and characterization of BADH cDNA from barley, a monocot, and the expression pattern of a BADH transcript. An open reading frame of 1515 bp encoded a protein which showed high homology to BADH enzymes present in other plants (spinach and sugar-beet) and in Escherichia coli. Transgenic tobacco plants harboring the clone expressed high levels of both BADH protein and its enzymatic activity. Northern blot analyses indicated that BADH mRNA levels increased almost 8-fold and 2-fold, respectively, in leaves and roots of barley plants grown in high-salt conditions, and that these levels decreased upon release of the stress, whereas they did not decrease under continuous salt stress. BADH transcripts also accumulate in response to water stress or drought, indicating a common response of the plant to osmotic changes that affect its water status. The addition of abscisic acid (ABA) to plants during growth also increased the levels of BADH transcripts dramatically, although the response was delayed when compared to that found for salt-stressed plants. Removal of plant roots before transferring the plants to high-salt conditions reduced only slightly the accumulation of BADH transcripts in the leaves.
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  • 61
    ISSN: 1573-5028
    Keywords: chloroplasts ; gene expression ; heat bleaching ; photosynthesis ; transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A shift in the ratio of chlorophyll (Chl) a and Chl b is an early indicator of heat bleaching in Euglena gracilis. This observation prompted us to consider whether or not changes in steady-state levels of chloroplast transcripts and in transcriptional activity could limit the synthesis of Chl a-binding proteins in bleaching plastids. We found that the mature transcripts for CP47 and CP43, the Chl a-binding apoproteins of the proximal antenna of photosystem II, decline sharply very early during bleaching. Our study also shows that transcription of psbB and psbC, the chloroplast genes encoding CP47 and CP43, remains essentially unchanged during the same interval. We conclude that posttranscriptional events, such as mRNA stability, could play a major role in initiating an irreversible loss of chloroplast function in Euglena at a moderately elevated temperature. Lack of these transcripts would eventually impair the assembly of photosystem II in thylakoids.
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  • 62
    ISSN: 1573-5028
    Keywords: gene expression ; gene family ; higher plants ; Hordeum vulgare ; metabolic regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have amplified and cloned DNA sequences derived from a gene encoding a SNF1 (sucrose-non-fermenting 1)-related protein kinase which differs from that previously reported from barley. Northern blot and polymerase chain reaction (PCR) analysis of RNA populations, using specific probes and oligonucleotide primers, indicated that the two SNF1-related genes are differentially regulated. One is expressed in all tissues, whereas the other is expressed at high levels in the seed endosperm and aleurone, but at levels undetectable by northern blot analysis in other tissues. Comparisons with other plant SNF1-related protein kinase genes suggest that the form which is expressed at greatly enhanced levels in the seed is less similar to the other plant homologues which have been reported and may be unique to cereals.
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  • 63
    ISSN: 1573-5028
    Keywords: gene expression ; light regulation ; nitrate reductase ; phytochrome ; signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have explored the possible involvement of the phosphoinositide (PI) cycle and protein kinase C (PKC) in the phytochrome (Pfr)-mediated light signal transduction pathway using nitrate reductase (NR) and phytochrome-I (PhyI) genes as model systems. We have shown earlier that phorbol myristate acetate (PMA) completely replaces the red light effect in stimulating nitrate reductase activity and transcript levels in maize. In this paper, we present detailed evidence to show that PMA mimics the red light effect and follows similar kinetics to enhance NR steady-state transcript accumulation in a nitrate-dependent manner. We also show that PMA inhibits phyI steady-state transcript accumulation in a manner similar to red light, indicating that a PKC-type enzyme(s) may be involved in mediating the light effect in both cases. Serotonin or 5-hydroxytryptamine (5-HT), a stimulator of PI turnover, was also found to mimic the red light effect in enhancing NR transcript levels and inhibiting phyI transcript accumulation, indicating the role of the PI cycle in generating second messengers for regulating the two genes. These results indicate that phytochrome-mediated light regulation of NR and phyI gene expression may involve certain common steps in the signal transduction pathway such as the PI cycle and protein phosphorylation by a PKC-type enzyme.
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  • 64
    ISSN: 1573-5028
    Keywords: auxin ; DNA binding factor ; gene expression ; glutathione S-transferase ; Nicotiana tabacum ; signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have analysed the promoter regions of two closely related auxin-regulated glutathione S-transferase genes. All active deletion constructs tested showed expression of the reporter gene β-glucuronidase (gusA) in root tips of young seedlings and newly developing lateral roots. Auxin treatment greatly enhanced the level of expression. The Nt103-1 promoter region −370/−276 was found to be necessary, at least as a quantitative element to confer auxin-responsiveness to a reporter gene, and sequences responsible for the auxin-responsiveness must be located downstream of −370. The region −651/−370 contains sequence information necessary for uninduced expression. The Nt103-35 promoter manifested its auxin-responsiveness within the −504/−310 region. Electrophoretic mobility shift analysis, using nuclear extracts from tobacco leaves and suspension cells, identified a factor binding to a sequence (ap103, TGAGTCT) at position −560 of the Nt103-1 promoter, which shows homology to the mammalian AP-1 site. A second factor was found to bind a sequence (as103, ATAGCTAAGTGCTTACG) with homology to the CaMV 35S promoter as-1 element. The as103 element is present in both promoters and positioned around −360, so within the region determined to be indispensable for the response to auxin. A third factor was found binding to the −276/−190 region of both promoters. Combined, these data point to the relevance of a 90 bp region for auxin-induced activity of both tobacco genes. The ASF-1 like factor binding to the as103 element within this region might be involved in mediating the auxin response.
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  • 65
    ISSN: 1573-5028
    Keywords: differential screening ; gene expression ; Lycopersicon esculentum ; rin ; ripening inhibitor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A tomato fruit cDNA library was differentially screened to identify mRNAs present at higher levels in fruit of the tomato ripening mutant rin (ripening inhibitor). Complete sequencing of a unique clone ERT D1 revealed an open reading frame with homology to several glutamate decarboxylases. The deduced polypeptide sequence has 80% overall amino acid sequence similarity to a Petunia hybrida glutamate decarboxylase (petGAD) which carries a calmodulin-binding site at its carboxyl terminus and ERT D1 appears to have a similar domain. ERT D1 mRNA levels peaked at the first visible sign of fruit colour change during normal tomato ripening and then declined, whereas in fruit of the ripening impaired mutant, rin, accumulation of this mRNA continued until at least 14 days after the onset of ripening. This mRNA was present at much lower levels in other tissues, such as leaves, roots and stem, and was not increased by wounding. Possible roles for GAD, and its product γ-aminobutyric acid (GABA) in fruit, are discussed.
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  • 66
    ISSN: 1573-5028
    Keywords: carotenoids ; cleavage site ; gene expression ; melon ; phytoene synthase ; ripening
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    Notes: Abstract A cDNA clone (MEL5), encoding a protein homologous to phytoene synthase (PSY), has been isolated from a climacteric melon fruit cDNA library, using the tomato cDNA clone TOM5 [34] as a heterologous probe. MEL5 hybridised to a transcript of 1.65 kb which suggested that the 1.36 kb clone, isolated originally, was not full-length. The missing 5′ end was isolated by a reverse transcriptase-polymerase chain reaction (RT-PCR)-based method. This enabled the full sequence of the protein to be deduced and the cleavage site of the transit peptide for chromoplast import to be predicted. Northern analysis of RNA extracted from fruit samples of different ripening stages as well as from roots, leaves and flower petals was used to examine the expression pattern of the corresponding mRNA. The transcript corresponding to MEL5 is present at low quantities in unripe (green) fruit, reaches its highest levels when the fruit turns from green to orange and persists at lower levels during later ripening stages. A similar transcript was also detected in flower petals and in trace amounts in leaves and roots. Genomic Southern analysis indicates that the clone is homologous to a low-copy-number gene family. Sequence analysis showed a high degree of conservation among plant PSYs.
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  • 67
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    Plant molecular biology 29 (1995), S. 11-23 
    ISSN: 1573-5028
    Keywords: drought ; flooding ; freezing tolerance ; gene expression ; salt
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    Topics: Biology
    Notes: Abstract Six cDNAs representing unique cold-induced sequences have been cloned from the hardy citrus relative Poncirus trifoliata. Among these, pBCORc115 and pBCORc119 were found to belong to the same gene family. Sequencing data indicated that pBCORc115 and pBCORc119 each contained an open reading frame, coding for a 19.8 kDa protein (COR19) and a smaller 11.4 kDa protein (COR11) respectively. Inspection of the deduced amino acid sequences revealed three large repeats in COR19, but only one was present in the COR11. Two elements: a Q-clustered tract and a K-rich motif were identified in each repeat. The K-rich motifs were similar to those of cotton D-11 and Group 2 LEA proteins. A Serine-cluster, a common feature in many Group 2 LEA-like proteins, was also found in these proteins, but it was in an unusual position at the carboxy-terminus. A bipartite motif of basic residues, similar to known nuclear targeting sequences, was also present in COR19 and COR11, suggesting that members of this protein family may have a nuclear targeting function. The expression of COR19 mRNA in response to cold acclimation, drought, flooding, and salinization was examined. COR19 expression in leaf tissue was induced in response to cold acclimation, but repressed during drought and flooding stress.
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  • 68
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    Plant molecular biology 29 (1995), S. 367-377 
    ISSN: 1573-5028
    Keywords: Avena sativa ; gene expression ; PHYA ; light regulation ; mRNA degradation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Gene-preferential oligonucleotide probes were used to determined the relative abundance and half-lives of distinct oat phytochrome A (PHYA) mRNAs. Oat PHYA mRNAs are highly conserved in the 5′-untranslated region and the coding region, but the 3′-untranslated region has an overall lower sequence conservation and was the source of gene-preferential probes. PHYA3 mRNA was estimated to be ca. 61% of the oat PHYA mRNA pool present in poly(A)+ RNA from dark-grown seedlings. The half-lives for PHYA3 and PHYA4 mRNAs were both estimated to be ca. 30 min, and a similar short half-life was estimated for the average PHYA mRNA. Sequence comparisons of PHYA mRNAs from four grass species identified conserved sequences within the 5′- and 3′-untranslated regions that might be important for PHYA mRNA degradation.
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  • 69
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    Keywords: Ascorbate peroxidase ; Arabidopsis thaliana ; gene expression ; guaiacol peroxidase ; ozone ; sulfur dioxide
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    Notes: Abstract The effects of ozone or sulfur dioxide on antioxidant enzymes were investigated in Arabidopsis thaliana. Plants were fumigated with 0.1–0.15 ppm ozone or sulfur dioxide up to about 1 week in an environment-controlled chamber. Both pollutants increased the activities of ascorbate peroxidase and guaiacol per-oxidase in leaves, but had little effect on the activities of superoxide dismutase, catalase, monodehydroascorbate reductase, dehydroascorbate reductase or glutathione reductase. Ozone was more effective than sulfur dioxide in increasing the activities of the peroxidases. Ascorbate peroxidase activity increased 1.8-fold without a lag period during fumigation with 0.1 ppm ozone, while guaiacol peroxidase activity increased 4.4-fold with a 1-day lag. Expression of the APX1 gene encoding cytosolic ascorbate peroxidase was further investigated. Its protein levels in leaves exposed to 0.1 ppm ozone for 4 or 8 days were 1.5-fold higher than in controls. Both ozone and sulfur dioxide elevated APX1 mRNA levels in leaves at 4 and 7 days, whereas at 1 day only ozone was effective. The induction of APX1 mRNA levels by ozone (3.4- to 4.1-fold) was more prominent than that by sulfur dioxide (1.6-to 2.6-fold). The APX1 mRNA level increased by day and decreased by night. Exposure of plants to 0.1 ppm ozone enhanced the APX1 mRNA level within 3 h, which showed a diurnal rhythm similar to that of the control. These results demonstrate that near-ambient concentrations of ozone as well as similar concentrations of sulfur dioxide can induce APX1 gene expression in A. thaliana.
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  • 70
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    Plant molecular biology 29 (1995), S. 823-831 
    ISSN: 1573-5028
    Keywords: afterripening ; aldose reductase ; Avena fatua ; gene expression ; LEA ; seed dormancy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To analyze the patterns of gene expression associated with seed dormancy in wild oat (Avena fatua), we have isolated cDNA clones corresponding to genes that are differentially expressed in dormant and afterripened line M73 embryos. Gene transcripts of these clones were maintained in embryos of imbibed dormant caryopses, but declined rapidly in afterripened embryos after imbibition. GA3 treatment of dormant caryopses, which breaks dormancy, could lower the transcript levels in dormant embryos. When the germination of afterripened caryopses was inhibited by high temperature (35 °C), the decline in abundance of the transcripts in afterripened embryos was arrested. These genes were expressed to various degrees in water-stressed, but not in unstressed, 7-day-old seedlings. The expression of the genes was also ABA-inducible in afterripened embryos. The expression patterns in non-dormant line SH430 wild oat were similar to those of afterripened M73. DNA sequence analyses indicated that some of the cDNA clones encode LEA (late embryogenesis-abundant) proteins and aldose reductase. The significance of the expression of these genes in maintaining seed dormancy or longevity is discussed.
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  • 71
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    Keywords: carbon catabolite repression ; cDNA ; gene expression ; stress-induced genes ; glucose-starvation ; Zea mays
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    Topics: Biology
    Notes: Abstract In order to isolate glucose-starvation-related cDNAs in maize (Zea mays L.) root tips, a cDNA library was constructed with poly(A)+ mRNA from 24 h starved root tips. After differential screening of the library, we isolated six different cDNAs (named pZSS2 and pZSS7) which were expressed during glucose starvation. Time course analysis revealed that maximum expression of five of these genes occurs 30 h after the onset of the starvation treatment. On the contrary, the expression of mRNAs corresponding to pZSS4 was maximal at an early stage of starvation and then dramatically decreased. The expression of this gene did not seem to be specific for glucose starvation. The pattern of induction of the genes corresponding to pZSS2, pZSS3, pZSS5, pZSS6 and pZSS7 revealed that non-metabolizable sugars such as L-glucose and mannitol induce mRNA transcription similarly to glucose starvation. When D-glucose or any other metabolizable sugar was supplied, the level of transcripts was reduced. Nucleotide sequence analyses of the six cDNAs allowed identification of five of them by comparison with sequence data bases. The protein encoded by clone pZSS2 is analogous to a wound-induced protein from barley. Clones pZSS4 to pZSS7 encode, respectively, a transmembrane protein, a cysteine protease, a metallothionein-like protein and a chymotrypsin/subtilisin-like protease inhibitor. Clone pZSS3 shares no significant homology with any known sequence.
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  • 72
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    Plant molecular biology 28 (1995), S. 647-656 
    ISSN: 1573-5028
    Keywords: abscission ; gene expression ; polygalacturonase ; ethylene ; auxin
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Abscission, organ separation, is accompanied by cell wall breakdown in separation layer cells. In tomato (Lycopersicon esculentum), ethylene-induced abscission is correlated with an increase in polygalacturonase (PG) and endo-β-1,4-D-glucanase (cellulase) activity. We have identified a putative, abscission-specific cDNA clone for PG, pTAPG1. The TAPG1 cDNA has 43% identity at the amino acid level with the tomato fruit PG. Genomic blot analysis suggests that the gene for TAPG1 is a member of a small subfamily of PG genes that is distinct from the tomato fruit PG. The TAPG1 cDNA hybridizes to mRNA expressed during the course of ethylene-induced leaf and flower abscission. A high level of PG transcript accumulation coincides with the occurrence of abscission. Auxin, an abscission inhibitor, and silver thiosulfate, an ethylene action inhibitor, suppressed accumulation of mRNA in leaf abscission zones complementary to the TAPG1 cDNA. Expression of TAPG1 transcripts is several-fold higher in flower abscission zones than in leaf abscission zones. The identification of cDNAs that encode abscission-specific PG provide and additional tool to study the regulation of abscission and cell wall dissolution in separation layer cells.
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  • 73
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    Plant molecular biology 28 (1995), S. 667-676 
    ISSN: 1573-5028
    Keywords: gene expression ; promoter ; glyceraldehyde-3-phosphate dehydrogenase ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transient expression experiments show that the maize GapA1 promoter exhibits a requirement for sequences contained within intron 1 and surrounding exon border regions for expression in maize Black Mexican Sweet cells. Maize GapA1-promoter constructs lacking intron 1 are inactive. Intron 1 and its exon border sequences, when reintroduced into constructs lacking introns, restore gene activity whereas intron 2 and its exon borders to not. The minimal promoter so defined encompasses roughly 250 bp upstream of the in vivo transcription start and appears also to include intron 1. An octameric sequence was identified in intron 1 of maize GapA1 which is similar to sequence motifs found in other maize introns known to increase transient expression. Partial restoration of gene expression in GapA1 constructs lacking intron 1 was achieved through insertion of the identified octameric sequence.
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  • 74
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    Plant molecular biology 28 (1995), S. 739-750 
    ISSN: 1573-5028
    Keywords: alcohol dehydrogenase ; fermentation ; gene expression ; pollen ; pyruvate decarboxylase ; respiration ; tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We characterized the genes coding for the two dedicated enzymes of ethanolic fermentation, alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC), and show that they are functional in pollen. Two PDC-encoding genes were isolated, which displayed reciprocal regulation: PDC1 was anaerobically induced in leaves, whereas PDC2 mRNA was absent in leaves, but constitutively present in pollen. A flux through the ethanolic fermentation pathway could be measured in pollen under all tested environmental and developmental conditions. Surprisingly, the major factor influencing the rate of ethanol production was not oxygen availability, but the composition of the incubation medium. Under optimal conditions for pollen tube growth, approximately two-thirds of the carbon consumed was fermented, and ethanol accumulated into the surrounding medium to a concentration exceeding 100 mM.
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  • 75
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    Plant molecular biology 28 (1995), S. 811-820 
    ISSN: 1573-5028
    Keywords: β-oxidation ; gene expression ; fruit ripening ; Mangifera indica ; peroxisomes ; thiolase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fruit ripening is a complex, developmentally regulated process. A series of genes have been isolated from various ripening fruits encoding enzymes mainly involved in ethylene and cell wall metabolism. In order to aid our understanding of the molecular basis of this process in a tropical fruit, a cDNA library was prepared from ripe mango (Mangifera indica L. cv. Manila). By differential screening with RNA poly(A)+ from unripe and ripe mesocarp a number of cDNAs expressing only in ripe fruit have been isolated. This paper reports the characterization of one such cDNA (pTHMF 1) from M. indica which codes for a protein highly homologous to cucumber, rat and human peroxisomal thiolase (EC 2.3.1.16), the catalyst for the last step in the β-oxidation pathway. The cDNA for the peroxisomal mango thiolase is 1305 bp in length and codes for a protein of 432 amino acids with a predicted molecular mass of 45 532 Da. Mango thiolase is highly homologous to cucumber thiolase (80%), the only other plant thiolase whose cloning has been reported, and to rat and human thiolases (55% and 55% respectively). It is shown by northern analysis that during fruit ripening THMF 1 is up-regulated. A similar pattern of expression was detected in tomato fruit. Wounding and pathogen infection do not appear to affect THMF 1 expression. The possible involvement of thiolase in fatty acid metabolism during fruit ripening will be discussed. To our knowledge this is the first report cloning of a plant gene involved in fatty acid metabolism showing an induction during fruit ripening.
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  • 76
    ISSN: 1573-5028
    Keywords: pyruvate kinase ; plastid ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Clones encoding two different forms of plastid pyruvate kinase (PKp; EC 2.7.1.40) have been isolated from both castor and tobacco seed cDNA libraries. One form, designated PKpA, from castor was described in a previous report, and the tobacco homologue of PKpA has now been isolated. In addition, a second cDNA, designated PKpG, has been identified and sequenced in both species. Western blot analysis, using antibodies raised against protein overexpressed from these clones, indicates that they encode the two predominant polypeptides of plastid pyruvate kinase from developing castor endosperm. In castor, both PKpA and PKpG are encoded by single genes. In the allotetraploid Nicotiana tabacum, there are two copies of each, one derived from each of the progenitors of this species. The expression of the genes for PKpA and PKpG was examined in various tissues from both castor and tobacco. In castor, both forms are expressed in developing and germinating endosperm and in the root but neither is expressed in the leaf. In tobacco, both forms are expressed in developing seeds but in mature tissues, PKpA is most abundant in roots and PKpG in leaves.
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  • 77
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    Plant molecular biology 27 (1995), S. 327-338 
    ISSN: 1573-5028
    Keywords: gene expression ; multi-gene family ; phenylalanine ammonia-lyase ; phenylpropanoids ; promoters ; secondary metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phenylpropanoid derivatives are a complex class of secondary metabolites that have many important roles in plants during normal growth and in responses to environmental stress. Phenylalanine ammonialyase (PAL) catalyzes the first step in the biosynthesis of phenylpropanoids, and is usually encoded by a multi-gene family. Genomic clones for three Arabidopsis thaliana PAL genes containing the entire protein-coding region and upstream and downstream sequences have been obtained and completely sequenced. Two A. thaliana PAL genes (PAL1 and PAL2) are structurally similar to PAL genes that have been cloned from other plant species, with a single intron at a conserved position, and a long highly conserved second exon. Previously identified promoter motifs plus several additional sequence motifs were found in the promoter regions of PAL1 and PAL2. Expression of PAL1 and PAL2 is both qualitatively and quantitatively similar in different plant organs and under various inductive conditions. A third A. thaliana PAL gene, PAL3, differs significantly from PAL1 and PAL2 and other sequenced plant PAL genes. PAL3 contains an additional intron, and its deduced amino acid sequence is less homologous to other PAL proteins. The PAL3 promoter region lacks several sequence motifs conserved between A. thaliana PAL1 and PAL2, as well as motifs described in other genes involved in phenylpropanoid metabolism. A. thaliana PAL3 was expressed at very low levels under the conditions examined.
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  • 78
    ISSN: 1573-5028
    Keywords: cis factor ; gene expression ; promoter ; transgenic rice ; wheat histone H3
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Type I element (CCACGTCACCGATCCGCG) is a well-conserved regulatory element found in proximal promoter region of a certain class of plant histone genes, that is composed of two independent cis-acting elements of the hexamer (ACGTCA) and the reverse-oriented octamer (GATCCGCG) motifs. To investigate functional role(s) of the type I element in regulation of a wheat histone H3 gene (TH012) promoter activity in vivo, base substitution mutations were introduced into the element and activities of the mutated promoters were examined in cultured rice cells, and in regenerated roots and anther walls of transgenic rice plants by employing a GUS reporter system. Mutations of each or both of the hexamer and the octamer motifs caused a reduction in the promoter activity in protoplasts transfected transiently or stably transformed calli. The mutation of the octamer motif with or without the mutation of the hexamer motif caused a marked reduction of the promoter activity in the root meristem of transgenic rice although the mutation of the hexamer motif alone caused a weak reduction. In contrast to these results, no effect of the mutations of either the hexamer or the octamer motif was found in the anther wall in which replication-independent activity of the H3 promoter was observed. Our results suggested that the hexamer and the octamer motifs may play important role(s) in regulation of replication-dependent but not of replication-independent expression of the wheat histone H3 gene.
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  • 79
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    Plant molecular biology 27 (1995), S. 743-752 
    ISSN: 1573-5028
    Keywords: plant hormone ; gibberellic acid ; GA-responsive ; gene expression ; HCA ; hydrophobic cluster analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A multiple gene family of at least four members, related to a GA-stimulated transcript (GAST1) from tomato, was characterized in Arabidopsis thaliana by analysing four related cDNAs, named GASA1 to GASA4. The corresponding peptides display comparable structural features: (1) a putative signal peptide of 18 to 23 residues; (2) a highly divergent hydrophilic region of about 22 amino acids; (3) a conservative 60 amino acid C-terminal domain containing 12 cysteines. This organization has also bean shown in two related peptides from tomato, GAST1 found in shoots and RSI-1 found in early lateral roots. Southern blot hybridization patterns showed single-copy genes for all four members of the GASA family. Accumulation of the various transcripts, monitored by northern blot hybridization, indicated that the various genes are expressed differentially in plant organs. Specific mRNAs were mostly detected in flower buds and immature siliques in the case of GASA1, in siliques and dry seeds in the case of GASA2 and 3, and in growing roots and flower buds in the case of GASA4. At least two of the GASA genes are activated in GA-deficient mutant ga5, as early as 4 to 8 h after spraying with 50 μM GA3. The complex patterns of expression and regulation of the various genes suggest that the related peptides are involved in a developmental regulation process in Arabidopsis.
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  • 80
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    Plant molecular biology 27 (1995), S. 953-967 
    ISSN: 1573-5028
    Keywords: calcium-dependent protein kinase ; gene expression ; protein phosphorylation ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated, from a cDNA library constructed from rice coleoptiles, two sequences, OSCPK2 and OSCPK11, that encode for putative calcium-dependent protein kinase (CDPK) proteins. OSCPK2 and OSCPK11 cDNAs are related to SPK, another gene encoding a rice CDPK that is specifically expressed in developing seeds [20]. OSCPK2 and OSCPK11-predicted protein sequences are 533 and 542 amino acids (aa) long with a corresponding molecular mass of 59436 and 61079 Da respectively. Within their polypeptide chain, they all contain those conserved features that define a plant CDPK; kinase catalytic sequences are linked to a calmodulin-like regulatory domain through a junction region. The calmodulin-like regulatory domain of the predicted OSCPK2 protein contains 4 EF-hand calcium-binding sites while OSCPK11 has conserved just one canonical EF-hand motif. In addition, OSCPK2-and OSCPK11-predicted proteins contain, at their N-terminal region preceding the catalytic domain, a stretch of 80 or 74 residues highly rich in hydrophilic amino acids. Comparison of the NH2-terminal sequence of all three rice CDPKs so far identified (OSCPK2, OSCPK11 and SPK) indicates the presence of a conserved MGxxC(S/Q)xxT motif that may define a consensus signal for N-myristoylation. OSCPK2 and OSCPK11 proteins are both encoded by a single-copy gene and their polyadenylated transcripts are 2.4 and 3.5 kb long respectively. OSCPK2 and OSCPK11 mRNAs are equally abundant in rice roots and coleoptiles. A 12 h white light treatment of the coleoptiles reduces the amount of OSCPK2 mRNA with only a slight effect on the level of OSCPK11 transcript. With anoxic treatments, OSCPK2 mRNA level declined significantly and promptly while the amount of OSCPK11 transcript remained constant.
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  • 81
    ISSN: 1573-5028
    Keywords: gene expression ; cyanobacterium ; nitrite reductase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nitrite reductase (NiR) gene (nirA) has been isolated and sequenced from the filamentous, thermophilic non-N2-fixing cyanobacterium Phormidium laminosum. Putative promoter-like and Shine-Dalgarno sequences appear at the 5′ end of the 1533 bp long nir-coding region. The deduced amino acid sequence of NiR from P. laminosum corresponds to a 56 kDa polypeptide, a size identical to the molecular mass previously determined for the pure enzyme, and shows a high identity with amino acid sequences from ferredoxin-dependent NiR. This cyanobacterial NiR gene has been efficiently expressed in Escherichia coli DH5α from the E. coli lac promoter and probably from the P. laminosum NiR promoter.
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    Plant molecular biology 27 (1995), S. 1183-1188 
    ISSN: 1573-5028
    Keywords: differential screening ; maize ; pith ; trpA ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA derived from an abundant maize pith mRNA transcript and its corresponding genomic equivalent have been isolated and characterized. High transcript levels are seen in the pith and young leaves of maize plants, while no transcript is detected in seed tissue of any age. The protein encoded by the isolated gene has considerable homology with tryptophan synthase alpha subunit (trpA) from other organisms and the cDNA clone can complement an E. coli trpA mutant. These data support the conclusion that this cDNA and the corresponding genomic clone encode a maize trpA protein.
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    Plant molecular biology 29 (1995), S. 91-98 
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; gene expression ; hypersensitive response ; oxidative stress ; ozone ; pathogenesis-related proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have used a reverse transcriptase polymerase chain reaction procedure (differential display) to isolate cDNAs corresponding to transcripts that accumulate in ozone-treated Arabidopsis thaliana. In this report we describe the characterization of an ozone-induced transcript, AtOZI1. AtOZI1 mRNA in untreated plants was detected at low levels in cotyledons, leaves, and flower buds and at higher levels in roots and mature flowers. AtOZI1 mRNA accumulation was transiently induced in leaves 3- to 5-fold within the first 6 h of ozone treatment. AtOZI1 mRNA accumulation was also transiently induced 3- to 6-fold by photopathogenic Pseudomonas strains. Sequence analysis of AtOZI1 revealed that it encodes a 8.6 kDa basic protein that contains a putative signal peptide and two potential phosphorylation sites. Our results suggest that AtOZI1 represents a novel stress-related protein that accumulates in response to the production of active oxygen species.
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    Plant molecular biology 29 (1995), S. 135-148 
    ISSN: 1573-5028
    Keywords: chromophyte ; Chrysophyceae ; light-harvesting complex protein gene ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated a gene from a library of nuclear DNA for a chlorophyll a/c-binding protein (named Cac for chl a/c by analogy with Cab for chl a/b) of a chromophyte alga, Giraudyopsis stellifer, and sequenced it. The comparison of the deduced amino acid sequence with other chl a/c-and chl a/b-binding protein sequences shows that structural and functional features, i.e. the arrangement ‘en X’ of the two A and B transmembrane helices and the putative chl a-binding sites, are shared by both Chlorophyta and Chromophyta. Moreover, in contrast to Chlorophyta, a very strong identity is found among Chromophyta in the C helix, suggesting a major function associated to this specific region. Nevertheless, the primary structure of the apoprotein does not seem affected by the pigment composition in Chromophyta. As in the few other examples currently known, we confirm that the cac genes are nuclear-encoded and are part of a multigenic family. Northern blots, performed on poly(A)+ mRNA from G. stellifer, give evidence that the cac gene is light-induced at a transcriptional level and that no expression can be observed in the dark.
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  • 85
    ISSN: 1573-5028
    Keywords: cathepsin B ; gene expression ; Nicotiana ; thiol protease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sequence analysis of a 1.33 kb clone from a root cDNA library of Nicotiana rustica revealed an open reading frame encoding a protein of 356 amino acids. The deduced protein has high levels of homology to human cathepsin B protease and a cathepsin B-like cysteine protease from wheat but much lower levels of homology with other plant cysteine proteinases. Southern blotting experiments suggest a limited number of cathepsin B-like genes are present in the genome of N. rustica and also that of N. tabacum. RNA analysis involving a range of tissues, harvested from both Nicotiana species 4–5 h after the beginning of a 16 h photoperiod, revealed the cathepsin B-like gene was being expressed strongly in roots, stem and developing flowers but weakly in mature leaves. Further analysis of RNA extracted from leaf tissue of N. tabacum revealed the gene showed rhythmic expression and also that its expression increased in response to wounding. Analysis of leaf tissues harvested during the latter part of a 16 h photoperiod (11 and 16 h after illumination commenced) showed that transcript levels were two three times higher than in leaf tissue harvested either towards the end of the dark period or 5 h after illumination commenced. When leaf tissue was wounded at 11:00 (5 h after plants were illuminated), and harvested for RNA extraction 6 h later, the level of cathepsin B-like transcript in mesophyll tissue was found to be increased ca. 2-fold relative to the level detected in unwounded controls.
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  • 86
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    Keywords: Arabidopsis thaliana ; chloroplast ; gene expression ; ω-3 fatty acid desaturase ; promoter ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Arabidopsis FAD7 gene encodes a chloroplast ω-3 fatty acid desaturase that catalyzes the desaturation of lipid-linked dienoic fatty acids (18:2 and 16:2). An 825 bp FAD7 promoter fragment upstream from the transcriptional start point contained several short sequences which were homologous to the cis-elements (box II, G-box, etc.) conserved in many light-responsive genes. We introduced the FAD7 promoter fused to the β-glucuronidase (GUS) or the luciferase (LUC) reporter gene into tobacco plants. The −825 promoter sequence conferred tissue-specific and light-responsive expression to both these reporter genes in transgenic tobacco, indicating that these expressions of the FAD7 gene were regulated mainly at the transcriptional level. Histochemical GUS staining showed that the activity of the FAD7 promoter is restricted to the tissues with chloroplast-containing cells although the staining was noticeably absent in the chloroplast-containing cells associated with vascular systems. The 5′ deletion experiments of the promoter revealed that the −362/ −166 region, containing two putative box II sequences, was responsible for the tissue-specific and light-responsive expression of the FAD7 gene.
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  • 87
    ISSN: 1573-5028
    Keywords: tomato ; polygalacturonase ; pectin methylesterase ; heat stress ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of extended heat stress on polygalacturonase (PG; EC 3.2.1.15) and pectin methylesterase (PME; EC 3.1.1.11) gene expression at mRNA, protein and activity levels in ripening tomato fruits were investigated. Steady state levels of PG mRNA declined at temperatures of 27°C and above, and a marked reduction in PG protein and activity was observed at temperatures of 32°C and above. Exogenous ethylene treatment did not reverse heat stress-induced inhibition of PG gene expression. Transfer of heat-stressed fruits to 20°C partly restored PG mRNA accumulation, but the rate of PG mRNA accumulation declined exponentially with duration of heat stress. Heat stress-induced inhibition of PME mRNA accumulation was recoverable even after 14 days of heat stress. In fruits held at 34°C, both PG and PME protein and activity continued to accumulate for about 4 days, but thereafter PG protein and activity declined while little change was observed in PME protein and activity. In spite of increases in mRNA levels of both PG and PME during the recovery of heat-stressed fruit at 20°C, levels of PG protein and activity declined in fruits heat-stressed for four or more days while PME protein and activity levels remained unchanged. Collectively, these data suggest that PG gene expression is being gradually and irreversibly shut off during heat stress, while PME gene expression is much less sensitive to heat stress.
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  • 88
    ISSN: 1573-5028
    Keywords: gene family ; gene expression ; photoreceptor ; phytochrome ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Data presented here define five tomato phytochrome genes (PHY) and indicate the existence of additional PHY in the tomato genome. Portions of each gene, encoding amino acids 203 through 315 in a consensus amino acid sequence, were amplified by polymerase chain reaction. Four of these genes, PHYA, PHYB1, PHYB2 and PHYE, are members of previously identified PHY subfamilies, while the fifth, PHYF, is identified as a member of a new PHY subfamily. PHYA, PHYB1, PHYB2 and PHYE fragments encode amino acid sequences that share 88% to 98% sequence identity with their Arabidopsis counterparts. The PHYF fragment, however, encodes a polypeptide that shares only 65% to 74% sequence identity with previously identified Arabidopsis phytochromes. A phylogenetic analysis suggests that PHYF arose soon after, or perhaps prior to, the origin of angiosperms. This analysis leads to the prediction that PHYF might be widespread among angiosperms, including both monocotyledons and dicotyledons. Each of the five tomato PHY is expressed as a transcript of sufficient size to encode a full-length phytochrome apoprotein. Two PHYF transcripts, 4.4 and 4.7 kb in length, have been detected in 9-day-old light-grown seedlings, consistent with either multiple transcription start sites or differential processing. Analyses of genomic Southern blots hybridized with radiolabelled RNA probes derived from the five tomato PHY, as well as Arabidopsis PHYC, indicate that the tomato genome contains as many as 9 to 13 PHY. The tomato PHY family is apparently not only different from, but also larger than, the PHY family presently described for Arabidopsis.
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  • 89
    ISSN: 1432-2242
    Keywords: 5S rDNA ; Evolution ; Protoplast fusion ; Somatic hybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleotide sequences of the 5S rRNA genes (5S rDNA) of two Solanum tuberosum breeding lines (R1 and B15) and of the Mexican wild species S. pinnatisectum were determined and compared with each other and to the 5S rDNA of other Solanaceae species (Lycopersicon esculentum, Nicotiana rustica and Petunia hybrida). The 5S rDNA repeats of the Solanum species are 324–329 bp in length, and they exhibit 91–95% sequence identity. Sequence variability is mainly located in a short region of the spacer separating the 5S rRNA coding regions. A synthetic 28-mer oligonucleotide constructed according to this region can be used as a specific hybridization probe to distinguish symmetric somatic hybrids between S. tubersosum breeding line B15 and S. pinnatisectum produced by protoplast fusion. Interestingly, the two Solanum breeding lines R1 and B15 differ also in this spacer region.
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  • 90
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    Theoretical and applied genetics 91 (1995), S. 189-194 
    ISSN: 1432-2242
    Keywords: Wheat ; HMW glutenin genes ; Polymerase chain reaction (PCR) ; Multigene families ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Specific amplification of the complete coding region of all six high-molecular-weight (HMW) glutenin genes present in hexaploid wheat was obtained by the polyerase chain reaction (PCR). Primers specific for the N-terminal region of the 1Dx gene and for the repetitive domain of the y-type HMW glutenin genes were also developed. Although the primers were constructed on the basis of the nucleotide sequences of HMW glutenin genes present in T. aestivum L. cv ‘Cheyenne’, they were very efficient in amplifying HMW glutenin genes of diploid and tetraploid wheat species. PCR analysis of HMW glutenin genes of T. urartu Tuman., T. longissimum (Schweinf. & Muschl.) Bowden and T. speltoides (Tausch) Gren. ex Richt, showed a high degree of length polymorphism, whereas a low degree of length variation was found in accessions of T. tauschii (Coss.) Schmal. Furthermore, using primers specific for the repetitive regions of HMW genes, we could demonstrate that the size variation observed was due to a different length of the central repetitive domain. The usefulness of the PCR-based approach to analyze the genetic polymorphism of HMW glutenin genes, to isolate new allelic variants, to estimate their molecular size and to verify the number of cysteine residues is discussed.
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  • 91
    ISSN: 1432-2242
    Keywords: Endosperm development ; Evolution ; Imprinting ; Incompatibility ; Reciprocal crosses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Endosperm Balance Number (EBN) and the polar-nuclei activation (PNA) hypotheses have been developed to interpret, explain and predict interspecific and interploidy crossabilities in the Solanums and the Gramineae, respectively. Although these two hypotheses evolved independently, they share a number of common features. Assignment of EBNs and ‘activation/response values’ (AVs/RVs) depend on plumpness, size, and germinability of hybrid seeds. Also, both hypotheses emphasize the importance of a balanced parental genic contribution for the normal development of endosperm. However, in the EBN hypothesis a 2 maternal∶1 paternal EBN ratio is a prerequisite for successful interspecific crossability, while the PNA hypothesis is based on the stimulative strength of the male nuclei to initiate mitotic divisions in the primary endosperm nucleus and is idependent on a 2∶1 ratio between the RV of the polar nuclei and the AV of the male gamete. Differences and similarities betweeen the EBN and PNA hypotheses are summarized and contrasted. It is proposed that EBN and PNA be considered as the same concept.
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  • 92
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    Theoretical and applied genetics 90 (1995), S. 356-363 
    ISSN: 1432-2242
    Keywords: Domestication ; Potato ; Chloroplast DNA ; Solanum stenotomum ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Five chloroplast DNA (ctDNA) types (W, T, C, S, and A) have previously been identified in the Andean tetraploid cultivated potatoes (Solanum tuberosum ssp. andigena) and three types (C, S, and A) in diploid cultivated potatoes (S. stenotomum). In this study, ctDNA types were determined for an additional 35 accessions of S. stenotomum and 97 accessions of putative ancestral wild species (15 of S. brevicaule, 26 of S. bukasovii, 4 of S. candolleanum, 25 of S. canasense, 17 of S. leptophyes, and 10 of S. multidissectum). The first five ctDNA types were also identified in S. stenotomum. The wild species were also polymorphic for ctDNA types except for S. brevicaule, which had only W-type ctDNA. T-type ctDNA was not found in any of the wild species and could have originated from W-type ctDNA after S. stenotomum arose. The other types of ctDNA evolved in wild species. The geographical distribution of each ctDNA type indicated that A-type ctDNA arose in central Peru and T-type ctDNA in the Bolivia-Argentine boundary. It is implied that potatoes were successively domesticated and that, in parallel, several wild species were differentiated from time to time and place to place from the ‘ancestral species’ complex. Subsequent sexual polyploidization formed a wide ctDNA diversity among the Andean tetraploid potatoes, and selection from them formed the limited ctDNA diversity found in Chilean tetraploid potatoes (ssp. tuberosum).
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  • 93
    ISSN: 1573-9368
    Keywords: human α1AT ; CAT ; transgenic mice ; gene expression ; liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The tissue-specific and developmental pattern of expression controlled by the proximal promoter (position −348 to+15) derived from the human α-1-antitrypsin (hα1AT) gene was studied in transgenic mice. The short promoter segment was linked to the chloramphenicol acetyltransferase (CAT) reporter gene. The transgene showed highly specific expression in the liver and the correct developmental pattern of regulation. Interestingly, this short promoter targets expression to the liver with a greater specificity than that reported for larger α1AT promoter fragments.
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  • 94
    ISSN: 1573-9368
    Keywords: gene expression ; transgenic mice ; thyroid hormone ; thyroid hormone response element ; muscle ; heart
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract During development in the murine ventricle, there is a switch in myosin heavy chain gene (MyHC) transcription. The β-MyHC is expressed in the ventricles during foetal development, but is shut down at or around birth, at which time α-MyHC transcription is activated. This antithetical switch is thought to be mediated by circulating levels of thyroid hormone (TH) and both low and high affinity thyroid response elements (TREs) have been identified in the proximal promoter region of the murine α-MyHC. Myosin gene expression in the atria is relatively unaffected by the TH status. Previously, we used site-directed mutagenesis of the promoter in a transgenic analysis to define those elements responsible for high levels of transcriptionin vivo. These analyses focused on the role(s) of twocis elements, TRE1 and TRE2 that are located at −129 to −149 and −102 to −120, respectively, on the α-MyHC promoter. Although the elements' ablation had differential effects on transgene expression, neither single mutation abolished transgene expression completely. Here, we show that mutating both elements results in a complete inactivation of the transgene in both ventricles and atria under euthyroid conditions. However, expression still can be detected in the hyperthyroid state, implying that, although the TRE1 and TRE2 elements are critical elements for high levels of α-MyHC transcriptionin vivo, other promoter sites can mediate at least some degree of transcriptional activation.
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  • 95
    ISSN: 1617-4623
    Keywords: Chlamydomonas reinhardtii ; Chloroplast aldolase ; Evolution ; Intron/exon structure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genomic clones encoding the plastidic fructose- 1,6-bisphosphate aldolase ofChlamydomonas reinhardtii were isolated and sequenced. The gene contains three introns which are located within the coding sequence for the mature protein. No introns are located within or near the sequence encoding the transit-peptide, in contrast to the genes for plastidic aldolases of higher plants. Neither the number nor the positions of the three introns of theC. reinhardtii aldolase gene are conserved in the plastidic or cytosolic aldolase genes of higher plants and animals. The 5′ border sequences of introns in the aldolase gene ofC. reinhardtii exhibit the conserved plant consensus sequence. The 3′ acceptor splice sites for introns 1 and 3 show much less similarity to the eukaryotic consensus sequences than do those of intron 2. The plastidic aldolase gene has two tandemly repeated CAAT box motifs in the promoter region. Genomic Southern blots indicate that the gene is encoded by a single locus in theC. reinhardtii genome.
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  • 96
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    Molecular genetics and genomics 249 (1995), S. 265-273 
    ISSN: 1617-4623
    Keywords: Agrobacterium rhizogenes ; Horizontal gene transfer ; Evolution ; Ri plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A tobacco homologue (trolC) of the rolC gene of the Agrobacterium rhizogenes Ri-plasmid was cloned and sequenced from Nicotiana tabacum L. cv. Havana 425. The coding region of trolC is similar in sequence (69–87% for DNA and 5489% for the deduced amino acid sequence) to rolC genes of the agropine, mannopine, and mikimopine strains of Ri-plasmids and the N. glauca rolC homologue. Southern analyses showed that trolC is encoded by a small gene family derived from the tomentosiformis ancestor of tobacco. This suggests that trolC resulted from an ancient transfer of DNA between A. rhizogenes and a progenitor of modern tobacco. Transcripts of trolC were detected in three morphologically distinct cultivars of tobacco. trolC mRNA accumulated in young leaves and shoot tips, but not in lower leaves and roots of mature plants. Accumulation of trolC mRNA in cultured leaf tissues was strongly down-regulated by auxin and induced by cytokinin. These results are of particular interest because they suggest that a gene of bacterial origin introduced during evolution can have a function in a modern plant.
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  • 97
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    Journal of plant research 108 (1995), S. 517-526 
    ISSN: 1618-0860
    Keywords: Cactaceae ; Evolution ; Water stress ; Wood ; Xeric adaptation ; Xylem
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Wide-band tracheids are a specialized tracheid type in which an annular or helical secondary wall projects deeply into the cell lumen. They are short, wide and spindle-shaped, and their bandlike secondary walls cover little of the primary wall, leaving most of it available for water diffusion. Wide-band tracheids appear to store and conduct water while preventing the spread of embolisms. They may be the most abundant tracheary element in the xylem, but they are always accompanied by at least a few vessels. Typically, fibers are absent wherever wide-band tracheids are present. Wide-band tracheids occur in the primary and secondary xylem of succulent stems, leaves and roots in genera of all three subfamilies of Cactaceae but were not found in the relictual genusPereskia, which lacks succulent tissues. In the large subfamily Cactoideae, wide-band tracheids occur only in derived members, and wide-band tracheids of North American Cactoideae are narrower and are aligned in a more orderly radial pattern than those of South American Cactoideae. Wide-band tracheids probably arose at least three times in Cactaceae.
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  • 98
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    International journal of anthropology 10 (1995), S. 177-181 
    ISSN: 1824-3096
    Keywords: Single cell gel electrophoresis assay ; Comet assay ; DNA damage ; DNA repair ; Human biology ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The estimation of genetic instability by direct extent of DNA damage and repair is an important aspect of studies on mutagenesis, carcinogenesis, aging and evolution. Different methods have been introduced from time to time in an effort to meet this need. Single cell gel electrophoresis (SCGE) assay is a new, simple and sensitive method of evaluating DNA damage and repair at individual cell level. This assay can be performed on extremely small number of cells and results can be obtained within a relatively short time. The SCGE assay has the potential to play an important role not only in the understanding of some of the fundamental aspects of human biology but also can be helpful in many practical ways.
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  • 99
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    Biology and philosophy 10 (1995), S. 129-180 
    ISSN: 1572-8404
    Keywords: Evolution ; explanation ; morality ; rationality ; normative communication ; fact/value distinction ; utilitarianism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract Gibbard's theory of rationality is “evolutionary” in terms of its result as well as its underpinning argument. The result is that judgments about what is “rational” are analyzed as being similar to judgments of morality — in view of what Darwin suggests concerning the latter. According to the Darwinian theory, moral judgments are based on sentiments which evolve to promote the survival and welfare of human societies. On Gibbard's theory, rationality judgments should be similarly regarded as expressing emotional attachments to behavioral norms which originate and function to coordinate social interaction. Consequently, Gibbard's theory of rationality might be used to illuminate Darwin's theory of morality, and vice versa. Additionally, as argued in the present essay, both can be further elaborated, and defended, by developing related themes in philosophical ethics: viz., connected with Hume and 20th-century emotivists. The main problem is that this general Darwinian approach faces widespread opposition nowadays, not only in ethics but in philosophy of science. The purpose of this essay is to analyze Gibbard's theory, critically and constructively, with emphasis on the pertinent commonalities in Darwin, Hume and the emotivists, while also critically addressing their common enemies. The pervasive methodological orientation is to relate this analysis to (philosophy of) science in general, and biological science in particular.
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  • 100
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    Biology and philosophy 10 (1995), S. 255-285 
    ISSN: 1572-8404
    Keywords: Animal ; Biology ; Consciousness ; Economics ; Evolution ; Natural Selection ; Suffering ; Welfare
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract Welfare biology is the study of living things and their environment with respect to their welfare (defined as net happiness, or enjoyment minus suffering). Despite difficulties of ascertaining and measuring welfare and relevancy to normative issues, welfare biology is a positive science. Evolutionary economics and population dynamics are used to help answer basic questions in welfare biology: Which species are affective sentients capable of welfare? Do they enjoy positive or negative welfare? Can their welfare be dramatically increased? Under plausible axioms, all conscious species are plastic and all plastic species are conscious (and, with a stronger axiom, capable of welfare). More complex niches favour the evolution of more rational species. Evolutionary economics also supports the common-sense view that individual sentients failing to survive to mate suffer negative welfare. A kind of God-made (or evolution-created) fairness between species is also unexpectedly found. The contrast between growth maximization (as may be favoured by natural selection), average welfare, and total welfare maximization is discussed. It is shown that welfare could be increased without even sacrificing numbers (at equilibrium). Since the long-term reduction in animal suffering depends on scientific advances, strict restrictions on animal experimentation may be counter-productive to animal welfare.
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