播娘蒿hmgr基因保守区片段的克隆与分析

Cloning and analysis of hmgr gene conserved fragments in Descurainia sophia

关键词:  播娘蒿  3-羟基-3-甲基戊二酰辅酶A还原酶  基因克隆

Key words:  Descurainia sophia  3-hydroxy-3-methyl-glutaryl-coenzyme A reductase(HMGR)  gene cloning

摘要:

通过比较6种植物的8条甲羟戊酸途径关键酶3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)基因同源区域,设计简并引物,利用RT-PCR技术成功地从播娘蒿叶中扩增出458bp的基因片段。通过BlastP比较,所推断的播娘蒿HMGR蛋白序列与拟南芥(NP_177775)、萝卜(CAA48610)、杜仲(AAV54051)、胡黄连(ABC74565)、喜树(AAB69726)、龙胆草(BAE92730)的一致性分别达到98%、96%、88%、89%、86%和87%。通过对蛋白质保守区、特征区以及进化树分析,证实该片段确为hmgr基因片段,该结果为首次报道。

Abstract:

Based on the design of degenerated oligonucleotides according to the conservative regions of eight 3-hydroxy-3-methylglutaryl-CoA reductases(HMGRs)from six plants and total RNA extracted from Descurainia sophia,a 458bp fragment of hmgr was obtained by using reverse transcription PCR strategy. Through sequence analysis by BlastP online,the resulting protein showed high homology to 3-hydroxy-3-methylglutaryl-CoA reductase,with 98% identification compared with Arabidopsis thaliana(NP_177775),96% with Raphanus sativus(CAA48610),88% with Eucommia ulmoides(AAV54051),89% with Picrorhiza kurrooa(ABC74565),86% with Camptotheca acuminata(AAB69726)and 87% with Gentiana lutea(BAE92730). Deduced amino acid sequences were also analyzed by PROSITE,ClustalX and MEGA Ver.3.1,and data present evidence for the existence of 3-hydroxy-3-methylglutaryl-CoA reductase in Descurainia sophia. This is the first report of the hmgr gene isolated from D.sophia.