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  • 1
    ISSN: 1432-0983
    Keywords: Chloroplast DNA ; Ribosomal protein and RNA polymerase genes ; Heterologous DNA: DNA hybridization ; Chlamydomonas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Considerable DNA sequence homology can be detected between the Escherichia coli genes coding for translational and transcriptional components and both the chloroplast and nuclear genomes of Chlamydomonas reinhardi. Labeled chloroplast DNA was demonstrated to hybridize to DNA fragments of the transducing phages λfus3 and λspc2 that encode ribosomal proteins of the α and S10 operons. Further, chloroplast DNA probes hybridize to fragments of λrtf d 18 that encode the β and β′ subunits of RNA polymerase. The regions homologous to the ribosomal protein and RNA polymerase genes were located on the chloroplast DNA physical map by probing restriction fragments of chloroplast DNA with phage or plasmid fragments carrying these E. coli genes. Probing nuclear DNA with bacterial gene probes revealed DNA fragments homologous to elongation factor and ribosomal protein genes. Most surprisingly, sequences homologous to the β subunit of RNA polymerase were found not only in chloroplast DNA but in nuclear DNA as well.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 30 (1996), S. 711-722 
    ISSN: 1573-5028
    Keywords: developmental regulation ; gene expression ; gene family ; phenylalanine ammonia-lyase ; tobacco ; wound response
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phenylalanine ammonia-lyase (PAL) catalyzes the first reaction in the general phenylpropanoid pathway leading to the production of phenolic compounds with a significant range of biological functions. A PAL gene we designated gPAL1, cloned from tobacco, consists of two exons separated by an intron of 1932 bp. Exon I, 398 bp, and exon II, 1747 bp, together encode a polypeptide of 715 amino acids. A putative TATA box and polyadenylation signal are found 144 bp upstream of the initiation codon and 193 bp downstream from the stop codon, respectively. Using various parts of gPAL1 as probes, genomic Southern blots indicated the presence of a small family of PAL genes in the tobacco genome that can be divided into two distinct subfamilies, one consisting of pal1 and pal2 and another of pal3 and pal4. Comparative genomic blot analysis of progenitor species (Nicotiana tomentosiformis and N. sylvestris) indicated that each species contains one PAL gene from each of the subfamilies, suggesting that pal1 and pal3 (or pal2 and pal4) diverged prior to the evolution of N. tabacum. Expression of the PAL gene family was examined using RNA gel blots. PAL transcript levels were significantly higher in flowers and roots than in leaves and stems of mature plants. PAL transcripts accumulate differentially during flower and leaf maturation in that mRNA levels decline during flower maturation but increase during leaf maturation. In leaves, PAL transcripts rapidly accumulated after wounding.
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  • 3
    ISSN: 1573-5028
    Keywords: ATP ; GTP ; protein kinase ; receptor ; rice ; signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A receptor-like protein kinase, OsPK10, has been cloned from rice (Oryza sativa). The 2.8 kb cDNA contains an open reading frame capable of encoding a peptide sequence of 824 amino acids. The topological features of the predicted OsPK10 protein include an N-terminal signal peptide, a cysteine-rich extracellular ligand-binding domain, a membrane-spanning segment, and a cytoplasmic domain possessing all the hallmarks of catalytic domains of eukaryotic protein kinases. The cytoplasmic domain was selectively expressed in Escherichia coli and assayed for kinase activity. The results show the protein is capable of autophosphorylation using either ATP or GTP as the phosphate donor. Phosphoamino acid analysis reveals phosphorylation of threonines, consistent with the substrate specificity indicated by sequence motifs in the catalytic core. A single amino acid substitution of Glu for Lys-528 completely abolishes autophosphorylation activity. DNA gel blot analyses suggest that the haploid rice genome contains a single copy of the OsPK10 gene. OsPK10 transcripts appear to be more abundant in shoots than in roots of rice seedlings.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-5028
    Keywords: Cab genes ; chlorophylla/b-binding protein ; Pisum sativum ; nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2048
    Keywords: Chlorophyll a/b protein ; Gene expression (Cab) ; Light and gene expression ; Light-harvesting complex (photosystem II) ; Multigene family (Cab) ; Pisum (Cab gene expression)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To measure transcript levels for individual members of the Cab (chlorophyll a/b protein) multigene family in pea under a range of developmental situations, we developed a system using cDNA synthesis, the polymerase chain reaction (PCR), and chemiluminescence detection. In order to design gene-specific PCR primers for all genes, a partial genomic clone for a fifth, Type I LHCII (light-harvesting complex of photosystem II) gene, Cab-9 The Cab-9 sequence appears in the Genbank/EMBL databases under the accession number M86906 , was isolated and sequenced. All seven known Cab genes in pea are expressed in light-grown buds and leaves, including several genes previously known only from genomic clones. There appear to be at least two groups of Cab genes in pea which differ in their response to light and development. The first group (consisting of Cab-8, AB96, Cab-215 and Cab-315) includes Type I, Type II and Type III genes, shows a relatively strong response to red light, and has bud transcript levels similar to or slightly higher than leaves. The second group, consisting of the Type I genes Cab-9, AB80 and AB66, shows little or no transcript accumulation 24 h after a red light pulse, and has higher transcript levels in leaves than in buds. Transcript levels for genes in this second group appear to be lower than those of the first group in all developmental situations examined. These data indicate that there has been an evolutionary divergence of the responses to light and development among the Type I LHCII genes.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 39 (1999), S. 231-242 
    ISSN: 1573-5028
    Keywords: protein kinase ; signal transduction ; photoreceptor ; gene regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The PsPK3 and PsPK5 genes of the garden pea encode protein-serine/threonine kinases whose catalytic domains are closely related to known signal transducing kinases from animals and fungi. The PsPK3 polypeptide is predicted to be located in the nucleus, whereas PsPK5 is a homologue of NPH1, the probable blue light receptor for phototropism from Arabidopsis. We found previously that when etiolated pea seedlings are illuminated with continuous white light, PsPK3 and PsPK5 transcript levels within apical buds decline substantially, reaching their minimum levels within one day of exposure to light. The role of light in regulating the expression of the PsPK3 and PsPK5 genes was investigated further. To gain insight into the rapidity with which expression changes, 6-day old, dark-grown pea seedlings were transferred to continuous white light, and PsPK3 and PsPK5 RNA levels monitored over the ensuing 24 h. While transcripts from the RbcS gene family increase, the PsPK3 and PsPK5 mRNAs decline rapidly to their minimum levels. PsPK5 mRNA declines 10-fold in ca. 2 h, whereas PsPK3 mRNA declines 4-fold in ca. 8 h. We used single pulses of light to elucidate which photoreceptor triggers the negative regulation of PsPK3 and PsPK5 gene expression. To assess phytochrome involvement, etiolated seedlings were treated with single pulses of red light, red followed by far-red light, or far-red light alone. RbcS induction by a red light pulse is reversible with a subsequent far-red light pulse, clearly showing that phytochrome mediates its induction. Likewise, RbcS expression is induced with a single pulse of blue light or a dichromatic pulse of red+blue light. However, none of these pulses trigger the PsPK3 and PsPK5 mRNA levels to decline. Given the lack of effectiveness of light pulses, etiolated seedlings were transferred to continuous light of three different qualities to determine the spectral sensitivity of PsPK3 and PsPK5 gene expression. Exposure to continuous red, continuous far-red, or continuous blue light causes the PsPK3 and PsPK5 mRNAs to decline and transcripts from the RbcS and Cab gene families to increase. One likely explanation is that phytochrome A mediates the responses of these genes to continuous far-red light. The effectiveness of continuous red light and blue light in triggering the reduction in PsPK3 and PsPK5 mRNA levels and the increase in RbcS and Cab mRNAs may imply the participation of additional phytochromes and/or cryptochromes. Thus, the PsPK3 and PsPK5 genes exhibit responsiveness to continuous light, but a lack of responsiveness to single light pulses that is unusual, and perhaps unique, among light-regulated genes.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 8 (1987), S. 53-59 
    ISSN: 1573-5028
    Keywords: DNA sequence ; ferredoxin ; genomic organisation ; pea ; phytochrome ; Fd: ferrdoxin protein ; Fed: ferredoxin gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have sequenced the pea (cv. Alaska) cDNA clone pEA46 (16) and shown that it codes for ferredoxin I. This clone has previously been shown to correspond to a transcript whose levels are controlled by phytochrome (Kaufman et al. (1985) Plant Physiol. 78: 388–393; Thompson et al. (1983) Planta 158: 487–500). The deduced amino acid sequence includes part of an hydrophobic transit sequence that shows only very limited homology to that of Silene pratensis ferredoxin. Genomic blotting analysis indicates that ferredoxin I is encoded by one or two genes. A genomic clone (4601) has been isolated that contains the ferredoxin gene and at least 14 kb of flanking sequences.
    Type of Medium: Electronic Resource
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