中药材2016，Vol.39Issue(12)：2706-2710,5.DOI:10.13863/j.issn1001-4454.2016.12.008
川泽泻鲨烯合酶基因的原核表达研究
Cloning and Prokaryotic Expression of Squalene Synthase(SS) from Alisma orientale
摘要
Abstract
Objectives:Construction of expression plasmid of SS and analysis of prokaryotic expression.Methods:SS coding sesquence was cloned from the fresh leaves of Alisma orientale by RT-PCR,and subsequently inserted into pET28 (with His tag) and pGEX-6t-1 (with GST tag),to construct two prokaryotic expression plasmids.Then the expression plasmids were transformed into Escherichia coli BL21 (DE3)competent cells and recombinant expression was explored with different induction temperatures and IPTG concentrations.Results:The SS protein was expressed as the inclusion body with the vector of pET28.The GST tag fusion with pGEX-6t-1 did not improve the solubility of SS.In addition,the highest protein expression of SS was observed at the induction temperature of 30 ℃ and there was no significant difference with induction by different concentrations of IPTG,indicating that the optimal induction temperature of SS was 30 ℃ and the IPTG concentration had little influence on the expression.Conclusions:The present work will be helpful and lay the foundation for further investigation of SS biological functions and quality improvement of Alisma orientale.关键词
川泽泻/鲨烯合酶基因/原核表达/泽泻醇Key words
Alisma orientale (Sam.) Juzep./Squalene synthase/Prokaryotic expression/Alisol分类
医药卫生引用本文复制引用
李丽霞,付羽萍,谌琴琴,王强..川泽泻鲨烯合酶基因的原核表达研究[J].中药材,2016,39(12):2706-2710,5.基金项目
全国第四次中药资源普查(2012-H-101) （2012-H-101）
