[目的]克隆刺五加叶绿体的ATP合酶β亚基cDNA并对其进行生物信息学分析.[方法]根据已知物种叶绿体ATP合酶β亚基基因的序列,设计1对同源引物,通过RT-PCR扩增得到刺五加ATP合酶β亚基cDNA,并对其进行序列比对及结构预测分析.[结果] RT-PCR扩增获得了长1 099 bp的刺五加ATP合酶β亚基cDNA,该基因编码366个氨基酸.序列比对及结构预测分析表明,刺五加ATP合酶β亚基基因编码的氨基酸与水稻的同源性最高,达96.41％.其二级结构中含有171个α螺旋(alpha helix),占46.72％；53个延伸链( extended strand),占14.48％;27个β折叠(beta turn),占7.38％；115个无规则蜷曲(random coil),占31.42％.第262 ～271位氨基酸为ATP合酶β亚基的标志性位点.整个多肽链无明显的疏水区域,初步认定为亲水性蛋白.[结论]该试验克隆得到的ATP合酶β亚基基因为叶绿体ATP合酶β亚基基因,为研究刺五加能量代谢对植物次生代谢的影响及了解植物ATP合酶的结构与功能提供了必要的信息.%[Objective] This study aimed to clone and analyze the cDNA of ATPase β subunit gene from Eleutherococcus senticosus. [ Method] A pair of homologous primers was designed according to the chloroplast ATPase β subunit gene sequences of the known species; then the gene cDNA of E. Senticosus were amplified by RT-PCR and compared with that of the known species; its structure was predicted finally. [ Result] 1 099 bp of ATPase beta subunit cDNA of E. Senticosus which encodes 366 amino acids was amplified by RT-PCR. Sequence comparison and structure prediction showed that amino acids encoded by the ATPase beta subunit gene of E. Senticosus shared the highest homology, up to 96.41% with that of Oryza sativa. In the secondary structure, the protein contained 171 alpha helixes accounting for 46. 72% , 53 extended strands accounting forl4.48% , 27 beta sheets accounting for 7. 38% and 115 random coils which took up 31.42% . The amino acids 262 -271 were the symbolic site of ATPase β subunit. The whole peptide chain had no obvious hydrophobic region and was primarily confirmed as a hydrophilic protein. [ Conclusion ] The cNDA of ATPase β subunit gene cloned from E. Senticosus in this study will provide important reference for learning the effect of energy metabolism on secondary metabolism, structure and function of ATPase in E. Senticosus.
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