热休克蛋白HSP65嵌合表达系统的构建和表达  

Expression and construction of chimeric vector about HSP65

作　　者：雷清[1] 蒋琳[1] 马超[1] 刘鹏[1] 

机构地区：[1]兰州生物制品研究所,兰州730046

出　　处：《微生物学免疫学进展》2007年第3期24-26,共3页Progress In Microbiology and Immunology

基　　金：甘肃省自然基金资助项目(3ZS051-A25-094)

摘　　要：设计一条含(GlySer)3连接肽的引物,从BCG经PCR得到HSP65-Linker的基因片段。将该基因定向克隆入原核表达载体pET42b(+),用大肠杆菌BL21(DE3)转化,构建了pET42-LHSP65的嵌合表达载体。DNA测序正确后经IPTG诱导表达,得到可溶性目的蛋白,Western-blot也证实了此重组蛋白可与抗HSP65发生特异性免疫反应。此重组体的构建为进一步探索HSP65的免疫佐剂功效奠定了基础。A primer including(Gly4Ser)3was designed,The LHSP65 gene was amplified by PCR from BCG,then ligated into expression vector pET42b(+) directly,Which was transformed into E.coli BL21(DE3) and obtained pET42-LHSP65 expression vector.It was detected by sequencing DNA and induced with IPTG.The recombinant protein is soluble,Western blotting analysis showed that the protein can react with anti-HSP65 antibody specifically.This chimeric vector is established for researching HSP65 protein.

关 键 词：HSP65 连接肽 基因表达 

分 类 号：Q78[生物学—分子生物学]