罂粟生物碱合成关键酶STORR的基因克隆及其表达载体构建  

Gene Cloning and Expression Vector Construction of STORR

作　　者：陈芳 常瑜 魏玉杰 张兆萍 龚永福 苏毓杰 杨振华 CHEN Fang;CHANG Yu;WEI Yujie

机构地区：[1]甘肃省农业工程技术研究院,甘肃武威733006 [2]甘肃省特种药源植物种质创新与安全利用重点实验室,甘肃武威733006

出　　处：《智慧农业导刊》2023年第15期73-76,共4页JOURNAL OF SMART AGRICULTURE

基　　金：财政部和农业农村部国家现代农业产业技术体系(CARS-21);中央引导地方科技资金发展项目(YDZX20216200001390);甘肃省自然基金项目(22JR11RA279)。

摘　　要：为研究罂粟生物碱合成关键酶(STORR)在罂粟中的作用,该研究以罂粟叶片cDNA为模板,利用RT-PCR技术,克隆得到STORR基因,并对其序列进行测序分析,利用双酶切方法构建pCambia2301-KY表达载体。研究结果表明,成功克隆到STORR基因,通过生物信息学分析,该基因包含一个长度为2703 bp的完整开放阅读框,编码900个氨基酸,分子式为C4476H7097N1207O1328S46,理论分子质量为10.05 kD,理论等电点为6.45,不稳定指数为41.70,为不稳定蛋白。用KpnI酶切载体pCambia2301-KY线性化,与回收纯化产物进行重组反应,成功构建STORR基因表达载体,将表达载体成功转化农杆菌GV3101。该研究为后续进行STORR基因功能研究提供理论依据。In order to study the role of key enzyme of alkaloid biosynthesis(STORR)in opium poppy(also known as Papaver somniferum L.),STORR gene was cloned from cDNA of opium poppy leaves by RT-PCR technique,and its sequence was sequenced.pCambia2301-KY expression vector was constructed by double enzyme digestion.The results showed that the STORR gene was successfully cloned.Bioinformatics analysis showed that the gene contained a complete open reading frame of 2703 bp,encoding 900amino acids,the molecular formula was C4476H7097N1207O1328S46,the theoretical molecular weight was 10.05 kD,the theoretical isoelectric point was 6.45and the instability index was 41.70.It was an unstable protein.The expression vector of STORR gene was successfully constructed by linearizing the vector pCambia2301-KY with KpnI enzyme and reacting with the purified product.The expression vector was successfully transformed into Agrobacterium tumefaciens GV3101.This study provides a theoretical basis for the follow-up study of STORR gene function.

关 键 词：罂粟 STORR基因 基因克隆 过表达载体 生物碱 

分 类 号：S567.212[农业科学—中草药栽培]