杜仲法尼烯基焦磷酸合酶基因cDNA全长的克隆与序列分析

Gene Cloning and Sequence Analysis of Farnesyl Pyrophosphate Synthase Genes from Eucommia ulmoides

采用RT-PCR法,从杜仲幼嫩叶片中分离法尼烯基焦磷酸基因c DNA全长序列,克隆并对该基因全长序列进行生物信息学分析。从杜仲嫩叶中共获得两个基因分别命名为Eu FPPs1和Eu FPPs2,测序结果表明两基因开放阅读框分别为1 047 bp和1 029 bp,推测分别可以编码348个和342个氨基酸残基,在线预测表明两个蛋白序列均存在两个聚丙烯合酶位点。系统进化分析结果表明,Eu FPPs1和Eu FPPs2分别聚集于不同的分支,它们之间的进化距离为0.352,但是在亲缘关系上均与楤木和人参最近,进化距离分别为0.281、0.287,0.175、0.184。

The farnesyl pyrophosphate synthase gene cDNA was isolated from leaves of Eucommia ulmoide by method of reveres transcription polymerase chain reaction （RT-PCR）, the gene was cloned and then to analyze the nucleotide sequence by bioinformatics software. Two genes was isolated from leaves of Eucommia ulmoide and named after EuFPPsl and EuFPPs2 respectively. The result of sequencing showed that the nu- cleotide sequences of EuFPPs 1 and EuFPPs2, respectively, contains an open reading frame of 1 047 bp and 1 029 bp encoding 348 amino acids and 342 amino acids. According to the online prediction, EuFPPs 1 and EuFPPs2 all contained two sites of polypropylene oxide synthase. The results of phylogenetie analysis sug- gested that EuFPPs 1 and EuFPPs2 gathered in different branches respectively, the evolutionary distance be- tween the EuFPPsl and EuFPPs2 were 0.352. While the closest genetic relatives of EuFPPsl and EuFPPs2 were the Aralia elata and Panax ginseng, the evolutionary distance were 0.281 and 0.287, 0.175 and 0.184, respectively.