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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 11 (1986), S. 171-176 
    ISSN: 1432-0983
    Keywords: Plastocyanin ; Photosynthesis ; Genomic and cDNA clones ; Sequence analysis ; Transcript ; Spinach
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plastocyanin is a member of photosynthetic electron transport chains that transfers electrons from cytochrome f to the oxidized P700 chlorophyll a pigment of the photosystem I reaction center. We have isolated and characterized cDNA- and genomic clones from spinach (Spinacia oleracea) encoding the complete plastocyanin-precursor polypeptide. The amino acid sequence derived from the nucleotide sequence shows that the precursor consists of 168 amino acid residues including a transit sequence of 69 residues. The precursor polypeptide has a predicted Mr of 16,917, the mature protein of 10,413. The available data indicate that plastocyanin derives probably from a single-copy gene. The coding region contains no intron. The size of the mRNA as determined by S1 nuclease protection experiments is approximately 660 nucleotides, although analysis of different cDNA clones suggests that longer RNA species do exist, approaching the size of the mRNA (850 bases) estimated by Northern blot techniques.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: Photosynthesis ; Ferredoxin-NADP+ oxidoreductase ; cDNA Nucleotide sequence ; Transit peptide ; Spinach
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In this paper, we report the structural characterization of several spinach ferredoxin-NADP+ oxidoreductase (FNR) cDNAs ranging in size from 0.9 to 1.5 kilobases. A comparison of the deduced amino acid sequence with the known amino acid sequence determined for the spinach protein establishes that 1.4–1.5 kpb inserts span the full length of the mature protein (314 amino acid residues; Mr = 35,382). These also include an N-terminal 55 amino acid transit peptide as well as maximally 171 and 214 nucleotide 5′ and 3′ untranslated sequences, respectively. Evidence has been obtained that various forms of FNR arise from at least two similar genes. The FNR precursor (369 amino acid residues) has a calculated molecular mass of 41.2 kDa. Comparison of the transit peptide with transit peptides from two other stromal proteins shows little similarity at the level of primary sequence but some common features in secondary structure predictions.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0983
    Keywords: Photosynthesis ; Photosystem I ; Subunits VI and VII ; Nucleotide sequence ; Spinach
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Recombinant phage which encode the entire precursor polypeptide for subunit VI of the photosystem I reaction center have been selected from a lambda gt11 cDNA expression library made from polyadenylated RNA of spinach seedlings. The sequence predicts a precursor polypeptide of 144 amino acids (Mr = 15.3 kDa), a mature protein of 95 residues (Mr = 10.4 kDa) that lacks methionine, histidine and cysteine, and a transit peptide of 49 residues (Mr = 4.9 kDa). The corresponding gene(s) is (are) designated psaH. The gene for subunit VII, psaC, has been located in the small single-copy region of the spinach plastid chromosome using a synthetic oligonucleotide and a heterologous hybridization probe. It is part of a polycistronic transcription unit that is constitutively expressed and processed. Putative processing products include a monocistronic RNA for psaC. The polypeptide chain of 81 (deduced) amino acids is highly conserved and strikingly resembles bacterial-type ferredoxins. It harbours cysteine residues that appear to be involved in the ligation of the two 4Fe4S centres A and B in photosystem I. None of the two subunits appears to be membrane-spanning, and subunit VI, as subunit VII, is located at the reducing (stromal) side of the reaction center. All available information on the major subunits of photosystem I from spinach has been combined into a (revised) topographic model. Evidence that the innermost — plastome-encoded — core of photosystem I represents an old bacterial heritage in present day chloroplasts is discussed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2242
    Keywords: Oenothera ; Chloroplast DNA fragments ; Reciprocal digestion ; Southern transfers ; Chloroplast rRNA hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1) More than 50 cleavage sites produced by the restriction endonucleases Sal I, Pst I, Kpn I, Sma I and Eco RI have been physically mapped on the 47 μm circular DNA molecule of the Euoenothera plastome IV. This plastome (= plastid genome) is considered to be the phylogenetically oldest of the subsection. 2) The DNA molecule is segmentally organized into four regions represented by a large duplicated sequence in inverted orientation whose copies are separated by two single-copy segments. The single-copy regions comprise about 14 and 57 Md in size, respectively. 3) The size of the inverted repeat, about 15 Md, was determined by restriction site mapping, by mapping of genes for ribosomal RNAs and by hybridization of a cRNA transcribed from a homologous part of Spinacia oleracea chloroplast DNA which appears to be phylogenetically conserved. 4) Hybridization of radio-iodinated spinach 16S, 23S and 5S chloroplast rRNA species to Southern blots of restricted plastome IV DNA has localized the rDNA to the inverted repeat regions, in the order given. The genes for 16S and 23S rRNA are separated by a 2.4 kbp spacer. 5) The physical map of the plastome IV DNA serves as basis for comparison with the DNA from the four other, closely related Euoenothera plastomes.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2242
    Keywords: Oenothera ; Evolution of plastid DNA ; Comparative restriction site mapping ; rDNA spacer differences ; Insertions and deletions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1) DNA has been isolated from the five genetically distinguishable plastid types of Oenothera, subsection Euoenothera. DNA of plastomes I to IV was obtained from plants with identical nuclear backgrounds containing the genotype AA of Oenothera hookeri whereas the DNA of plastome V came from Oenothera argillicola (genotype CC). 2) The DNAs of the five basic Euoenothera wild-type plastomes can be distinguished by restriction endonuclease analysis with Sal I, Pst I, Kpn I, Eco RI and Bam HI. The fragment patterns exhibit distinct common features as well as some degree of variability. 3) Physical maps for the circular DNAs of plastome I, II, III and V could be constructed using the previously detailed map of plastome IV DNA (Gordon et al. 1981). This has been achieved by comparing the cleavage products generated by restriction endonucleases Sal I, Pst I and Kpn I which collectively result in 36 sites in each of the five plastome DNAs, and by hybridization of radioactively labelled chloroplast rRNA or chloroplast cRNA probes of spinach to Southern blots of appropriate restriction digests. The data show that the overall fragment order is the same for all five plastome DNAs. Each DNA molecule is segmentally organized into four regions represented by a large duplicated sequence in inverted orientation whose copies are separated by two single-copy segments. 4) The alterations in position of restriction sites among the Euoenothera plastome DNAs result primarily from insertions/deletions. Eleven size differences of individual fragments in the Sal I, Pst I and Kpn I patterns measuring 0.1–0.8 Md (150–1,200 bp) relative to plastome IV DNA have been located. Most changes were found in the larger of the two single-copy regions of the five plastomes. Changes in the duplication are always found in both copies. This suggests the existence of an editing mechanism that, in natural populations, equalizes or transposes any change in one copy of the repeat to the equivalent site of the other copy. 5) Detailed mapping of the two rDNA regions of the five plastomes, using the restriction endonucleases Eco RI and Bam HI which each recognize more than 60 cleavage sites per DNA molecule, disclosed a 0.3 Md deletion in plastome III DNA and a 0.1 Md insertion in plastome V DNA relative to DNA of plastome IV, I and II. These changes are most probably located in the spacer between the genes for 16S and 23S rRNA and are found in both rDNA units.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1617-4623
    Keywords: Photosynthesis ; Rieske iron-sulfur precursor protein ; cDNA nucleotide sequence ; Transit peptide ; Spinach
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Summary Several cDNA clones encoding the entire Rieske FeS-precursor protein of the chloroplast cytochrome b 6 f-complex have been isolated by high density plaque immunoscreening of a phage lambda gt11 cDNA expression library, made from poly A+-RNA of spinach seedlings. The identity of the cDNAs has been confirmed by N-terminal amino acid sequencing of the purified protein. The nucleotide sequence indicates a protein of 247 amino acid residues including a putative transit sequence of 68 amino acids corresponding to molecular masses of 26.3 kDa (precursor) and 18.8 kDa (mature protein; 179 amino acid residues). Alignteins of the sequence with sequences from Rieske FeS-proteins of respiratory electron transport chains, two of bacterial and three of mitochondrial origin, shows little sequence homology, but remarkable similarity in secondary structure including a putative N-terminal transmembrane segment of about 25 residues and the peptides CTHLGCV and CPCHGS in the C-terminal region of the protein that are involved in the binding of the Fe2S2-cluster.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1617-4623
    Keywords: Gene expression ; Light regulation ; Photosynthesis ; Plastid signal
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nuclear-encoded genes for proteins of the photosynthetic maschinery represent a particular subset of genes. Their expression is cooperatively stimulated by discrete factors including the developmental stage of plastids and light. We have analyzed in transgenic tobacco the plastid- and light-dependent expression of a series of 5′ promoter deletions of various nuclear genes from spinach, of fusions of defined promoter segments with the 90-bp 35S RNA CaMV minimal promoter, as well as with mutations in sequences with homologies to characterizedcis-elements, to address the question of whether the plastid signal and light operate via the same or differentcis-acting elements. In none of the 160 different transgenic lines (representing 32 promoter constructs from seven genes) analyzed, could significant differences be identified in the responses to the two regulatory pathways. The data are compatible with the idea that both signals control the expression of nuclear genes for plastid proteins via the samecis-acting elements.
    Type of Medium: Electronic Resource
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