丝瓜籽核糖体失活蛋白基因Luffin-a原核表达载体的构建  

Construction of prokaryotic expression vector for ribosome inactivating protein gene Luffin-a of Luffa seeds

作　　者：孙晓东[1] 王燕[1] 罗超[1] 王珺[1] 桑明[1,2] 

机构地区：[1]湖北医药学院基础医学研究所,湖北十堰442000 [2]武汉大学动物实验中心/ABSL-3Lab,湖北武汉430071

出　　处：《中国医药导报》2013年第7期22-24,共3页China Medical Herald

基　　金：湖北省十堰市科学技术研究与开发项目(项目编号:十科发2010-047S;十科发[2009]29S43);湖北医药学院研究生启动金项目(项目编号:2009QDJ06;2005QDJ08)

摘　　要：目的探讨丝瓜籽核糖体失活蛋白基因Luffin-a原核表达载体的构建方法。方法从Pubmed中查到Luffin-a的mRNA全序列,用RT-PCR的方法从未成熟的丝瓜种子中克隆Luffin-a基因,T-A克隆后鉴定核苷酸序列,构建原核表达载体pET-15b(+)-Luffin-a,转化大肠杆菌BL21(DE3),并经IPTG诱导表达;SDS-PAGE鉴定Luffin-a表达产物。结果克隆出Luffin-a基因,IPTG诱导后大肠杆菌培养液和超声破碎的菌体沉淀中都有Luffin-a表达产物。结论本研究成功构建了Luffin-a原核表达载体。Objective To investigate the method of construction of prokaryotic expression vector for ribosome inactivating protein gene Luffin-a of Luffa seeds. Methods Whole mRNA sequence of Luffin-a was found from Pubmed database, Luffin-a gene from immature Luffa seeds was cloned by RT-PCR method, and the sequence of nucleotides was identified after T-A clone, then the prokaryotic expression vector of pET-15 b （＋）-Luffin-a was constructed, the vec tor was transfected into E. coli BL21 （DE3）, and the expression was inducted by IPTG. Luffin-a expression product was identified by SDS-page. Results Luffin-a gene was cloned. Luffin-a expression product was detected in the supernatant of E. coli culture solution and ultrasonic broken bacteria after IPTG induction. Conclusion This study successfully constructs prokaryotic expression vector of Luffin-a.

关 键 词：丝瓜籽 核糖体失活蛋白 Luflfin—a基因 

分 类 号：R3[医药卫生—基础医学]