论文部分内容阅读
[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1099bp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] The research aimed to clone cDNA of ATPase β subunit of Acanthopanax senticosus chloroplast and analyze its bioinformatics. [Method] According to the sequence of chloroplast ATP synthase β subunit gene of a known species, a pair of homologous primers was designed and the cDNA of Acanthamoeba ATP synthase β subunit was amplified by RT-PCR and sequenced And structure prediction analysis. [Result] The cDNA of Acanthopanax senticosus ATP synthase β subunit with a length of 1099 bp was obtained by RT-PCR. The gene encoded 366 amino acids. Sequence alignment and structure prediction showed that the amino acids encoded by Acanthopanax senticosus ATP synthase β subunit gene had the highest homology with rice (96.41%). The secondary structure contains 171 alpha helixes (46.72%), 53 extended strands (14.48%), 27 β-turns (7.38%), and 115 random strands (random coil), accounting for 31.42%. The amino acids 262-271 are indicative of the ATP synthase β subunit. No obvious hydrophobic region of the polypeptide chain, initially identified as hydrophilic protein. [Conclusion] The ATP synthase β subunit gene cloned by this experiment was the chloroplast ATP synthase β subunit gene. To study the effect of energy metabolism of Acanthopanax senticosus on plant secondary metabolism and understand the structure and function of plant ATP synthase The necessary information.