pBV-BPI600重组表达载体的构建及其在E.coli中的表达  

Construction of pBV-BPI600 Recombinant Expression Vector and its Expression in E.coli

作　　者：安云庆[1] 刘箐 柯岩[1] 沈海中[1] 

机构地区：[1]首都医科大学微生物学和免疫学教研室

出　　处：《首都医科大学学报》2001年第1期1-5,共5页Journal of Capital Medical University

基　　金：国家自然科学基金!资助项目 (395 70 6 6 6 )

摘　　要：①采用RT PCR技术 ,以HL 60细胞mRNA为模板扩增获得编码BPI氨基端 1 93个氨基酸 (rBPI2 1)的基因片段 (BPI60 0 ) ;EcoRI/BamHI酶切扩增产物获得BPI 2 0 0bp和BPI 4 0 0bp 2个基因片段。②成功构建PUC1 8 BPI2 0 0和PUC1 8 BPI4 0 0重组克隆载体 ,DNA测序分析结果与文献报道一致。③成功构建pBV2 2 0 BPI60 0重组表达载体 ;转化E .coliDH5α感受态细胞 ,诱导目的重组蛋白表达 ;经SDS PAGE电泳和Western blot鉴定 ,证实表达的重组蛋白确为外源基因 (BPI 60 0bp)编码的产物BPI2 1重组蛋白。BPI 600 bp gene which encode 193 amino acid fragments (rBPI 21 ) BPI N terminal was amplified by RT PCR from mRNA that were extracted from HL 60; It was digested by EcoRI and BamHI to obtain BPI 400 bp and BPI 200 bp fragments.②PUC18 BPI200 and PUC18 BPI400 recombinant cloning vector were successfully constructed, and sequences were identical with those of the report. ③pBV BPI600 recombinant expression vector was successfully constructed, It was transformated into the competent E.coli DH5α and BPI 21 recombinant protein was expressed by temperature induced method. The results of SDS PAGE and Western blot has proved that the expressed recombinant protein is rBPI 21 which is encoded by BPI600 gene.

关 键 词：重组杀菌 渗透增强蛋白 pBV220表达载体 PUC18克隆载体 

分 类 号：Q782[生物学—分子生物学] R392.11[医药卫生—免疫学]