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  • 1
    ISSN: 1432-1432
    Keywords: Balbiani rings ; Nucleotide sequence ; 3′ ends ; Repeat units ; Evolutionary conservation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An interspecies comparison was made between the 3′ ends of Balbiani ring genes fromChironomus. The comparison was focused on the BR2.2 gene, and a part at the 3′ end fromChironomus pallidivittatus (which included also a segment of the gene core) was cloned. Its sequence, and other previously published BR sequences from this species and fromChironomus tentans were used in the analysis. The 3′ parts of these repetitive genes can be divided into a region belonging to the core of the genes followed by a terminal region. In the core region the repeats (each of which consists of a constant part and a subrepeated part) are highly similar and the constant parts show little interspecies differentiation. Furthermore, the two parts of the repeats are units in an evolutionary and probably also functional sense. The terminal region contains modified constant units, usually isolated betwen acidic so-called cys regions, the whole arrangement lying upstream of an intron toward a 3′-terminal exon. Most of the modified constant units are mosaics in rates of evolution with stable outer quarters bordering to equally stable cys regions and a central half with a very high rate of evolution. One of the terminal units, present only in the BR2.2 gene and second from the end, differs distinctly not only from corresponding core units but also from other terminal units in the three normally active BR genes. It lies upstream of all cys regions and is evolutionarily conserved over most of its length. Furthermore, two-dimensional protein structure prediction does not exclude an endoproteolytic cleavage site in this unit. Such a site appears unlikely in other terminal or core regions. This is of interest in view of evidence for intracellular cleavage of the BR2.2 terminal region with liberation of a part containing a DNA-binding domain (Botella et al. 1988). All in all the fine anatomy of evolutionary changes at the BR gene termini shows interesting correlations with postulated functional relations and may have predictive value in the further functional analysis of this part of the gene.
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  • 2
    ISSN: 1432-1432
    Keywords: Balbiani rings ; cDNA clones ; Nucleotide sequence ; Repeat units ; Subrepeats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A new type of repeat unit was isolated from Balbiani ring 1 ofChironomus pallidivittatus and designated, BR1β repeat. It consists of a constant and a subrepeated part, like previously described units belonging to the core blocks of the BR genes. The subrepeated part contains 10-codon subrepeats with an arrangement similar to the subrepeats of the previously described BR2β gene. The present unit differs from earlier reported core units firstly in a much lower number of copies (about 15) per genome, which are tandemly arranged. Secondly, the number of subrepeats per BR1β repeat unit can show great variations. On the basis of the pattern of codon usage, three types of subrepeats can be distinguished. One type lies 5′-proximal in the subrepeat array and consists of variable numbers of subrepeats almost identical at the nucleotide level. The last complete subrepeat represents another type, with consistent differences in codon usage as compared to subrepeats of the proximal type. Finally, there is an intermediate type represented by the subrepeat preceding the distal one. Here, codon characteristics from proximal and distal subrepeats are mixed in a patchy and irregular way. The evolution of the arrays can be understood either as being the result of subrepeat formation in two steps (occurring before and after amplification of whole repeat units) or as the result of a continuous process in which there is evidence for participation of gene conversion.
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  • 3
    ISSN: 1573-5028
    Keywords: calcium ; cycloheximide ; indole-3-acetic acid (IAA) ; mechanical strain ; protein kinase ; salt stress ; touch ; Vigna radiata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protein kinases are important in eukaryotic signal transduction pathways. In this study we designed degenerate oligonucleotides corresponding to two conserved regions of protein kinases and using the polymerase chain reaction (PCR) have amplified a 141 bp fragment of DNA from mungbeans (Vigna radiata Rwilcz cv. Berken). Sequence analysis of the PCR products indicates that they encode several putative protein kinases with respect to their identity with other known plant protein kinases. Using one of the six fragments (CPK3-8), we isolated a 2022 bp cDNA (VrCDPK-1) from a Vigna radiata λgt11 library. VrCDPK-1 has a 96 bp 5′-untranslated region and a 465 bp 3′-untranslated region and an open reading frame of 1461 bp. VrCDPK-1 contains all of the conserved regions commonly found in calcium dependent protein kinases (CDPK). VrCDPK-1 shares 24 to 89% sequence identity with previously reported sequences for plant CDPKs at the protein level. southern analysis revealed the presence of several copies of the CDPK gene. VrCDPK-1 expression was stimulated when mungbean cuttings were treated with CaCl2, while treatment with MgCl2 had no effect. We are reporting for the first time a CDPK gene in mungbean which is inducible by mechanical strain. Cuttings treated with indole-3-acetic acid (IAA) or subjected to salt stress showed an increase in VrCDPK-1 expression. There was a dramatic stimulation in VrCDPK-1 expression 6 h after cuttings were treated with cycloheximide.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 24 (1994), S. 757-766 
    ISSN: 1573-5028
    Keywords: touch ; calcium ; indole-3-acetic acid ; salt stress ; light ; signal transduction ; Vigna radiata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two different calmodulin (CaM) cDNAs (MBCaM-1 and MBCaM-2) were isolated from a vigna radiata λgt 11 library by screening with a heterologous Arabidopsis cDNA probe (TCH-1). Both cDNAs are 85% homologous inside the coding region but are highly divergent outside this region. The polypeptides encoded by MBCaM-1 and MBCaM-2 are identical except for two conservative substitutions at positions 7 and 10. Southern analysis revealed that both cDNAs are encoded by different genes. Expression studies revealed different patterns of expression of both genes. MBCaM-1 mRNA exhibited a dramatic transient increase in response to touch, while MBCaM-2 expression showed a steady but small increase as compared to MBCaM-1. When plants were grown in complete darkness MBCaM-1 was undetectable and MBCaM-2 exhibited very low levels of expression. One hour after exposure of etiolated seedlings to light MBCaM-1 showed no change, while MBCaM-2 expression was increased. After a 6 h exposure to light there was an induction of both MBCaM-1 and MBCaM-2; however, the magnitude of this increase was much greater for MBCaM-2. When plants were grown under a 16 h light/8 h dark cycle the mRNA levels for MBCaM-1 were lower during the light period and increased during the beginning of the night cycle, while MBCaM-2 showed no change. Plants treated with indole-3-acetic acid had a peak in MBCaM-1 expression 6 h after treatment initiation with a slight decline 3 h after the peak, while MBCaM-2 showed a steady but small increase over time as compared to MBCaM-1. When plants were subjected to salt stress they showed an increase in MBCaM-1 expression 2 h after treatment initiation reaching a maximum after 4 h with no further increase after 6 h, while MBCaM-2 remained unchanged over the time course.
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