白细胞介素2-绿脓杆菌外毒素融合基因的克隆及高效表达  

Cloning and High Level Expression of Interleukin 2-pseudomonas Exotoxin Fusion Gene in Escherichia coli

作　　者：高基民[1] 黄洪莲[1] 王小宁[1] 徐舲飞 郑仲承[2] 刘新垣[2] 

机构地区：[1]第一军医大学免疫教研室,广州510515 [2]中国科学院上海生物化学研究所,上海200031

出　　处：《生物工程学报》1996年第2期129-133,共5页Chinese Journal of Biotechnology

基　　金："863";国家自然科学基金;全国青年科学基金;广东省青年科学基金

摘　　要：利用聚合酶链式反应和寡核苷酸介导的定向诱变技术构建了白细胞介素2-绿脓杆菌外毒素IL2-PE40、IL2-PE40KDEL、IL2-PE66^(4Glu)和IL2-PE66^(4Glu)KDEL融合基因的原核表达重组质粒,并实现了高效表达,表达产物占菌体总可溶蛋白的20％～30％。此外,由于这一表达质粒在IL-2cDNA与PE基因连接处引入了唯一的SmaⅠ位点,其5＇、3＇端分别含有唯一的EcoRⅠ、PstⅠ位点,因此可方便地用其它基因替换IL-2或PE基因而获得相应融合蛋白的表达质粒。Using PCR and oligonucleotide-directed mutagenesis, recombinant plasmids which express IL2-PE fusion genes, such as IL2-PE40, IL2-PE40KDEL, IL2-PE664Ght and IL2-PE664GlnKDEL, were constructed and expressed in E. colt at high level; 20-30 percent of total soluble bacterial proteins are the fusion protein. They exist as inclusion bodies in E. coli. In addition , there are unique EcoR I , Pst I and Sma I sites , respectively, in these expression vectors, so it will be conveniently changed to another expression vectors which contain other cytokines or toxins.

关 键 词：白细胞介素-2 绿脓杆菌外毒素 融合蛋白 制备 

分 类 号：R392－33[医药卫生—免疫学] R394.8[医药卫生—基础医学]