八角莲愈伤组织中开环异落叶松脂素脱氢酶基因克隆、表达及功能鉴定  

Cloning,expression and functional identification of secoisolariciresinol dehydrogenase gene from Dysosma versipellis callus

作　　者：沈云[1] 陈日道[1] 解可波 邹建华[1] 戴均贵[1] 

机构地区：[1]中国医学科学院北京协和医学院药物研究所天然药物活性物质与功能国家重点实验室,北京100050

出　　处：《中国中药杂志》2016年第24期4568-4571,共4页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81273405)

摘　　要：开环异落叶松脂素脱氢酶(secoisolariciresinol dehydrogenase,SDH)是鬼臼毒素(podophyllotoxin)生物合成途径中的关键酶。该研究通过基于SDH基因保守性序列的同源PCR方法,结合快速扩增c DNA末端(RACE)技术从八角莲Dysosma versipellis愈伤组织中克隆得到2个SDH候选基因SO282和SO1223,并实现了以上基因在大肠杆菌中的可溶性表达。体外酶活性分析表明重组SO282具有SDH活性,为后续研究八角莲中鬼臼毒素的生物合成奠定了基础。Secoisolariciresinol dehydrogenase （SDH） is a key enzyme involved in the biosynthetic pathway of podophyllotoxin. In this study, two SDH candidate genes,SO282 and SO1223, were cloned from callus of Dysosma versipellis by homology-based PCR and rapid amplification of cDNA end （RACE）. The SDH candidate genes were expressed in Escherichia coli and the subsequent enzyme as- say in vitro showed that recombinant SO282 had the SDH activity. These results pave the way to the follow-up investigation of the bio- synthetic of podophyllotoxin.

关 键 词：八角莲 开环异落叶松脂素脱氢酶 鬼臼毒素 生物合成 

分 类 号：S567.239[农业科学—中草药栽培]