The chloroplastchIL gene of the green algaChlorella vulgaris C-27 contains a self-splicing group I intron |
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Authors: | Meenu Kapoor Tatsuya Wakasugi Koichi Yoshinaga and Masahiro Sugiura |
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Institution: | (1) Center for Gene Research, Nagoya University, 464-01 Nagoya, Japan;(2) Department of Chemistry, Faculty of Science, Shizuoka University, 422 Shizuoka, Japan;(3) Present address: Department of Biology, Faculty of Science, Toyama University, Gofuku 3190, 930 Toyama, Japan |
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Abstract: | ThechiL gene product is involved in the light-independent synthesis of chlorophyll in photosynthetic bacteria, green algae and non-flowering plants. The chloroplast genome ofChlorella vulgaris strain C-27 contains the first example of a splitchiL gene, which is interrupted by a 951 bp group I intron in the coding region. In vitro synthesized pre-mRNA containing the entire intron and parts of the flanking exon sequences is able to efficiently self-splice in vitro in the presence of a divalent and a monovalent cation and GTP, to yield the ligated exons and other splicing intermediates characteristic of self-splicing group I introns. The 5 and 3 splice sites were confirmed by cDNA sequencing and the products of the splicing reaction were characterized by primer extension analysis. The absence of a significant ORF in the long P9 region (522 nt), separating the catalytic core from the 3 splice site, makes this intron different from the other known examples of group I introns. Guanosine-mediated attack at the 3 splice site and the presence of G-exchange reaction sites internal to the intron are some other properties demonstrated for the first time by an intron of a protein-coding plastid gene. |
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Keywords: | Self-splicing chiL Group I intron Chlorella Chloroplast |
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