日本血吸虫组织蛋白酶L1基因在大肠埃希菌中的表达  

Expression of Proteinase Cathepsin L1 Gene of Schistosoma japonicum in Escherichia coli

作　　者：易冰[1] 李卓雅[1] 何蔼[1] 郑小英[1] 郑斌[1] 周豪杰[1] 王轶[1] 张瑞林[1] 余南[1] 詹希美[1] 

机构地区：[1]中山大学中山医学院寄生虫学教研室,广州510080

出　　处：《中国寄生虫学与寄生虫病杂志》2004年第2期76-79,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.301708377)

摘　　要：目的　在原核表达体系大肠埃希菌中进行日本血吸虫组织蛋白酶L1(SjCL1)基因表达。　方法　通过PCR从质粒pcDNA3 SjCL1中扩增得到SjCL1基因 ,定向克隆至原核表达载体pGEX 4T 1中 ,构建重组质粒pGEX SjCL1,将该重组质粒转化大肠埃希菌JM10 9,转化子经异丙基 β D 硫代半乳糖苷诱导表达 ,采用十二烷基硫酸钠 聚丙烯酰胺凝胶电泳 (SDS PAGE)和蛋白质印迹法 (Westernblotting)分析SjCL1基因表达产物。　结果　获得长约 1kb的PCR片段 ,构建了pGEX SjCL1质粒。SDS PAGE和Westernblotting检测表达产物 ,相对分子质量为 62 0 0 0。Objective To express the proteinase cathepsin L1 gene of Schistosoma japonicum (SjCL1) in Escherichia coli JM109 cells. Methods The SjCL1 gene was amplified from the recombinant plasmid pcDNA3-SjCL1 by PCR. The gene was cloned into a prokaryotic expression vector pGEX-4T-1 to construct a recombinant plasmid pGEX-SjCL1. The E.coli JM109 cells were transformed with the recombinant plasmid pGEX-SjCL1 and the transformants were induced by IPTG to express the recombinant protein, the target protein was then identified by SDS-PAGE and Western blotting. Results A 1 kb length PCR product was obtained and a recombinant plasmid pGEX-SjCL1was constructed. The expression product was detected by SDS-PAGE and Western blotting and an expression band about 62 000 was found. Conclusion The SjCL1 gene is effectively expressed in the E. coli JM109 cells.

关 键 词：日本血吸虫 组织蛋白酶类 基因表达 大肠埃希菌 原核表达载体 

分 类 号：R383.24[医药卫生—医学寄生虫学]