Kapoor M, Wakasugi T, Yoshinaga K, Sugiura M
Center for Gene Research, Nagoya University, Japan.
Mol Gen Genet. 1996 Apr 10;250(6):655-64. doi: 10.1007/BF02172976.
The chlL gene product is involved in the light-independent synthesis of chlorophyll in photosynthetic bacteria, green algae and non-flowering plants. The chloroplast genome of Chlorella vulgaris strain C-27 contains the first example of a split chlL gene, which is interrupted by 951 bp group I intron in the coding region. In vitro synthesized pre-mRNA containing the entire intron and parts of the flanking exon sequence 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.
chlL基因产物参与光合细菌、绿藻和非开花植物中叶绿素的非光依赖合成。普通小球藻C-27株的叶绿体基因组包含一个断裂chlL基因的首个实例,其编码区被一个951 bp的I类内含子中断。在体外合成的包含整个内含子和部分侧翼外显子序列的前体mRNA,在存在二价阳离子、一价阳离子和GTP的情况下能够在体外高效自我剪接,产生连接的外显子和其他自我剪接I类内含子特有的剪接中间体。通过cDNA测序确认了5'和3'剪接位点,并通过引物延伸分析对剪接反应产物进行了表征。在将催化核心与3'剪接位点分开的长P9区域(522 nt)中不存在明显的开放阅读框,这使得该内含子不同于其他已知的I类内含子实例。鸟苷介导的对3'剪接位点的攻击以及内含子内部存在的G交换反应位点是首次由蛋白质编码质体基因的内含子证明的其他一些特性。