刺五加鲨烯合酶基因cDNA的克隆与序列分析  

Cloning and Sequence Analysis of Squalene Synthase Gene in Eleutherococcus senticosus

作　　者：龙月红[1] 邢朝斌[1] 王明艳[1] 吴鹏[1] 陈龙[1] 梁能松[1] 何闪[1] 

机构地区：[1]河北联合大学生命科学学院,唐山063000

出　　处：《生物技术通报》2012年第2期112-116,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30701086);河北省自然科学基金项目(C2009001252)

摘　　要：采用改良的异硫氰酸胍法提取刺五加总RNA,逆转录为cDNA,根据已报道的人参鲨烯合酶基因(squalene synthasegene,SS)cDNA序列设计引物,利用RT-PCR法克隆刺五加SS基因的cDNA序列。克隆得到长度为1258 bp的刺五加SS基因cDNA序列,开放阅读框全长1248 bp,编码415个氨基酸残基,GenBank登录号为HQ456918,与人参的SS1、SS2和SS3氨基酸序列一致性分别为91.73%、97.59%和96.63%。首次分离并报道了刺五加SS基因cDNA序列,为刺五加苷生物合成中关键酶的表达分析及调控机理研究提供参考。Totle RNA of E.senticosus was extracted using improved isothiocyanate method,and cDNA was obtained by reverse transcription. Primers was designed according to cDNA of Panax ginseng SS.SS sequence was colned by RT-PCR.The cloned cDNA sequence of E.senticosus SS was 1 258 bp,with ORF span 1248 bp,coding 415 amino acid.Login number of E.senticosus SS for GenBank was HQ456918.The homology with SS1,SS2 and SS3 of P.ginseng was 91.73%,97.59% and 96.63%,respectively.We first extracted and reported cDNA sequence of E.senticosus SS.This study made foundation for key enzyme expression and regulate mechanism analysis in eleutheroside biosynthesis pathway.

关 键 词：刺五加 鲨烯合酶基因 克隆 RT-PCR 

分 类 号：S567.19[农业科学—中草药栽培] Q943.2[农业科学—作物学]