柱花草编码CP12蛋白的cDNA克隆及其序列分析

Cloning and Sequence Analysis of CP12 Gene from Stylosanthes guianensis

叶绿体蛋白12(chloroplast protein 12,CP12)是一个存在于叶绿体与光合作用相关的蛋白。以亲本柱花草热研2号叶片为材料,根据豌豆CP12及柱花草响应低温的cDNASSH文库中CP12克隆片段设计引物,采用RT-PCR,从柱花草cDNA中克隆获得柱花草热研2号CP12基因全长序列,并在GenBank登陆(登录号:HQ906668),命名为SgCP12。柱花草热研2号CP12基因全长426bp,包含399 bp的ORF。通过生物信息学分析结果表明,CP12的C端具有高度保守的结构域。同源性分析表明,该基因与豌豆的CP12基因氨基酸序列同源性最高。

Chloroplast protein 12 （CP12） locates in chloroplast related to photosynthesis. Based on the sequence of a fragment of CP12 in a cDNA SSH library responsive to cold in Stylosanthes guianensis and the highly homologous regions from pea CP12 gene, a pair of primes were designed and used for cloning CP12 from Stylosanthes guianensis by using RT-PCR. A full-length cDNA of CPI2 gene was isolated from the leaves of Stylosanthes guianensis Reyan. 2 by application of RT-PCR techniques, and was named SgCP12 （HQ906668）. It was found to be 426hp long and contained an open reading frame （ORF） of 399 hp, encoding a peptide of 132 amino acids. Bioinformatic analysis showed that SgCPl2 was most homologous to that pea CP12, and its C-termlnal domain was highly conserved.