毒性小分子蜂毒肽基因的克隆修饰、融合载体构建及表达研究  

Modification and Fusion Vector Construction of Small Venomous Melittin Gene and Its Expression in E.coli

作　　者：潘凌子[1] 方宏筠[1] 李大力[1] 王关林[1] 

机构地区：[1]辽宁师范大学生命科学学院,辽宁大连116029

出　　处：《辽宁师范大学学报（自然科学版）》2006年第3期340-343,共4页Journal of Liaoning Normal University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30270488)

摘　　要：通过定点诱变的方法,对小分子蜂毒肽(melittin)基因进行修饰,引入了羟胺裂解位点,使蜂毒肽与其前体蛋白得以在体外融合表达.将融合的蜂毒肽前体蛋白(premelittin)经双酶切后克隆到原核表达载体PET-42a(+)中,PCR鉴定后转化至宿主菌BL21(DE3)中,在异丙基硫代--βD-半乳糖苷(IPTG)诱导下,在大肠杆菌中实现融合表达.为利用基因工程的方法表达毒性小分子多肽、实现该类药物的产业化打下基础.Melittin gene was modified by inserting hydroxylamine clearage site through site-directed mutagenesis and cloned into expression vector pET42a（＋）. After identified by PCR, the recombinant plasmid was transformed into E. coli BL21 （DE3）. The fusion protein was expressed in E. coli under IPTG inducing. The research laid the foundation for producing these small venomous peptides by using genetic engineering method, and made it more possible for industrialization of this kind medicines before long.

关 键 词：蜂毒肽 修饰 融合表达 

分 类 号：Q786[生物学—分子生物学]