麻疯树磷酸烯酮式丙酮酸羧化酶pepc基因全长cDNA克隆及序列分析

Cloning and Sequence Analysis of Full-length cDNA of pepc Gene from Jatropha curcas

应用RT-PCR和RACE法从麻疯树总RNA中分离出pepc基因全长cDNA,长度为3 142 bp,阅读框2 898 bp,编码965个氨基酸,在GenBank中登录,序列号为EU069413。它的氨基酸序列与蓖麻、陆地棉、橙、大豆、花生、烟草、油菜、拟南芥的氨基酸同源性分别为94.94%、92.46%、90.60%、90.50%、90.50%、88.33%、84.61%、82.44%。该基因编码了pepc基因家族中的pepc1,蛋白属于C3型PEPC。推测了pepc编码蛋白分子量为110.6 kD,并对其稳定性、二级结构、疏水性等特性进行了分析,最后确定了该蛋白的功能位点和结构域。

Applying RT-PCR and RACE method,3 142 bp pepc gene was isolated from Jatropha curcas.The sequence analysis showes that the pepc gene including an open reading frame is 2 898 bp in length,and encodes a protein of 965 amino acid.Its amino acid sequence shares 94.94%,92.46%,90.60%,90.50%,90.50%,88.33%,84.61%,82.44% identity with those of pepc from Ricinus communis,Gossypium hirsutum,Citrus sinensis,Glycine max,Arachis hypogaea,Nicotiana tabacum,Brassica napus,Arabidopsis thaliana,respectively.The pepc gene has been accepted by GenBank,and the accession number of gene sequence is EU069413.The deduced protein molecular weight of pepc is 110.6 kD,and belong to pepc1 in gene family and C3 form.Its stability,secondary structure,hydrophobicity profile were analysed,and the functional sites and characterized domains were confirmed.