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  • 1
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] A set of broad host range cloning vectors has been constructed from the IncW plasmid pSa. These vectors have been constructed from the transfer defective deletion derivative pSa151, which encodes resistance to kanamycin and spectinomycin–streptomycin. Two of the vectors also contain the ...
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  • 2
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Evidence is presented showing that the set of broad-range cloning pSa plasmids are stably maintained in Escherichia coli. Cells containing these plasmids are generally less fit in glucose-limited chemostat cultures than plasmid-free cells. However, the insertion of a λ cos-containing DNA ...
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  • 3
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 277 (1979), S. 324-325 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The E. coli plasmid pBRHSB is a 5.2 megadalton derivative of pBR316 (ref 2) which codes for ampicillin (Ap) and tetra-cycline (Tc) resistance. It was constructed by inserting a chemically synthesised EcoRl linker into the RNA polymerase binding site (that is, the promoter) of the Tc-resistance ...
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  • 4
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 293 (1981), S. 309-311 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Most bacterial resistance to chloramphenicol is due to a chloramphenicol acetyl transferase (CAT) encoded on R-factors in Gram-negative4 and Gram-positive bacteria5'6. In Gram-negative bacteria, the Cmr gene on the transposable genetic element TnP codes for a 219-amino acid protomer (EC 2.3.28). In ...
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 12 (1993), S. 293-297 
    ISSN: 1432-203X
    Keywords: Lycopersicon esculentum ; Genetic marker ; Intraspecific genetic map ; DNA polymorphism ; Isozyme ; RFLP ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The existing molecular genetic maps of the tomato, Lycopersicon spp, are constructed based on isozyme and RFLP polymorphisms between tomato species. These maps are useful for certain applications but have few markers that exhibit sufficient polymorphisms for intraspecific analysis and manipulations within the cultivated tomato. The purpose of this study was to investigate the relative potential of RAPD technology, as compared to isozymes and RFLPs, to generate polymorphic DNA markers within cultivated tomatoes. Sixteen isozymes and 25 RFLP clones that were known to detect polymorphism between L. esculentum and L. pennellii, and 313 random oligonucleotide primers were examined. None of the isozymes and only four of the RFLP clones (i.e., 16%) revealed polymorphism between the cultivated varieties whereas up to 63% of the RAPD primers detected one or more polymorphic DNA fragments between these varieties. All RAPD primers detected polymorphism between L. esculentum and L. pennellii genotypes. These results clearly indicate that RAPD technology can generate sufficient genetic markers exploiting sequence differences within cultivated tomatoes to facilitate construction of intraspecific genetic maps.
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  • 6
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract PCR with oligonucleotide primers that corresponded to two highly homologous regions, in terms of amino acid sequence, of plant peroxidases was used to amplify a specific DNA fragment from a mixture of rice (Oryza sativa L.) cDNAs. We then screened a cDNA library prepared from mRNAs of rice shoots utilizing the product of PCR as probe. Two cDNA clones, prxRPA and prxRPN, were isolated. They encode distinct isozymes of peroxidase. Sequence analysis indicated that the clones encode mature proteins of approximately 32 kDa, both of which possess a putative signal peptide. Comparison of the amino acid sequences of the two rice peroxidases showed that they are about 70% similar to each other but are only 40% to 50% similar to other plant peroxidases. RNA blot hybridization revealed that mRNAs that corresponded to prxRPA and prxRPN cDNAs accumulate at high levels in roots but only at low levels in stems and leaves. In various tissues of rice plants, levels of both mRNAs were stimulated by wounding and by ethephon. These results indicate that at least two isozymes of peroxidase are expressed not only in shoots but also in roots of rice plants, and that the expression of these genes is influenced by ethylene which is the simplest plant hormone.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 16 (1991), S. 797-805 
    ISSN: 1573-5028
    Keywords: alevrone ; embryo ; multigene family ; northern blotting ; intact germinating seeds
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Steady-state levels of mRNA from individual α-amylase genes were measured in the embryo and aleurone tissues of rice (Oryza sativa) and two varieties of barley (Hordeum vulgare L. cv. Himalaya and cv. Klages) during germination. Each member of the α-amylase multigene families of rice and barley was differentially expressed in each tissue. In rice, α-amylase genes displayed tissue-specific expression in which genes RAmy3B, RAmy3C, and RAmy3E were preferentially expressed in the aleurone layer, genes RAmy1A, RAmy1B and RAmy3D were expressed in both the embryo and aleurone, and genes RAmy3A and RAmy2A were not expressed in either tissue. Whenver two or more genes were expressed in any tissue, the rate of mRNA accumulation from each gene was unique. In contrast to rice, barley α-amylase gene expression was not tissue-specific. Messenger RNAs encoding low- and high-pI α-amylase isozymes were detectable in both the embryo and aleurone and accumulated at different rates in each tissue. In particular, peak levels of mRNA encoding high-pI α-amylases always preceded those encoding low-pI α-amylases. Two distinct differences in α-amylase gene expression were observed between the two barley varieties. levels of high-pI α-amylase mRNA peaked two days earlier in Klages embryos than in Himalaya embryos. Throughout six days of germination, Klages produced three times as much high-pI α-amylase mRNA and nearly four times as much low-pI α-amylase mRNA than the slower-germinating Himalaya variety.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 14 (1990), S. 655-668 
    ISSN: 1573-5028
    Keywords: gene duplication ; genomic clones ; molecular evolution ; nucleotide sequence ; regulatory DNA sequences
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To establish the size and organization of the rice α-amylase multigene family, we have isolated 30 α-amylase clones from three independent genomic libraries. Partial characterization of these clones indicates that they fall into 5 hybridization groups containing a total of 10 genes. Two clones belonging to the Group 3 hybridization class have more than one gene per cloned fragment. The nucleotide sequence of one clone from Group 1, λOSg2, was determined and compared to other known cereal α-amylase sequences revealing that λOSg2 is the genomic analog of the rice cDNA clone, pOS103. The rice α-amylase genes in Group 1 are analogous to the α-Amy1 genes in barley and wheat. λOSg2 contains sequence motifs common to most actively transcribed genes in plants. Two consensus sequences, TAACA G A A and TATCCAT, were found in the 5′ flanking regions of α-amylase genes of rice, barley and wheat. The former sequence may be specific to α-amylase gene while the latter sequence may be related to a ‘CATC’ box found in many plant genes. Another sequence called the pyrimidine box ( T C CTTTT T C ) was found in the α-amylase genes as well as other genes regulated by gibberellic acid (GA). Comparisons based on amino acid sequence alignment revealed that the multigene families in rice, barley and wheat shared a common ancestor which contained three introns. Some of the descendants of the progenitor α-amylase gene appear to have lost the middle intron while others maintain all three introns.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 16 (1991), S. 579-591 
    ISSN: 1573-5028
    Keywords: DNA rearrangement ; gene duplication ; genomic sequences ; Oryza sativa ; PCR ; quantitative primer extension
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rice genomic clones containing eight different α-amylase genes have been previously classified into five groups based on DNA hybridization studies and restriction site mapping. This report describes the clustering of three Group 3 genes (RAmy3A, RAmy3B and RAmy3C) within 28 kb of genomic DNA. The genes are separated from each other by about 5 kb and transcribed in the same direction. At the protein level, RAmy3B and RAmy3C are 95% homologous while each is 78% homologous to RAmy3A. All three genes have relatively small introns in the first and third positions. RAmy3A; however, has an additional 409 bp intron in the second intron insertion site. Nucleotide sequence comparisons of the coding and 3′ flanking regions suggest that clustering of the RAmy3 genes occurred by gene duplication resulting from unequal crossing-over at repetitive sequences. A comparison of the 5′ flanking regions revealed several sequences that may be involved in transcription. Expression of RAmy3B/C first appears in the germinating seed after two days and at a higher level after four days. Quantitative primer extension analysis indicates that RAmy3B and RAmy3C contribute 25% and 75%, respectively, of the transcripts from this cluster at four days of germination. No primer extension band specific to RAmy3A transcripts could be detected at this time point. However, RAmy3A PCR products could be amplified from RNA isolated from embryo-derived callus tissue.
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  • 10
    ISSN: 1573-5028
    Keywords: glucanase ; gene expression ; pathogenesis response ; stress response ; plant growth regulators ; gene family
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A rice β-glucanase gene was sequenced and its expression analyzed at the level of mRNA accumulation. This gene (Gns1) is expressed at relatively low levels in germinating seeds, shoots, leaves, panicles and callus, but it is expressed at higher levels in roots. Expression in the roots appears to be constitutive. Shoots expressGns1 at much higher levels when treated with ethylene, cytokinin, salicylic acid, and fungal elicitors derived from the pathogenSclerotium oryzae or from the non-pathogenSaccharomyces cereviseae. Shoots also expressGns1 at higher levels in response to wounding. Expression in the shoots is not significantly affected by auxin, gibberellic acid or abscisic acid. The β-glucanase shows 82% amino acid similarity to the barley 1,3;1,4-β-D-glucanases, and from hybridization studies it is the β-glucanase gene in the rice genome closest to the barley 1,3;1,4-β-glucanase EI gene. The mature peptide has a calculated molecular mass of 32 kDa. The gene has a large 3145 bp intron in the codon for the 25th amino acid of the signal peptide. The gene exhibits a very strong codon bias of 99% G+C in the third position of the codon in the mature peptide coding region, but only 61% G+C in the signal peptide region.
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