论文部分内容阅读
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。 结论 SjCL1基因在大肠埃希菌中以融合形式得到表达
Objective To express Schistosoma japonicum cathepsin L1 (SjCL1) gene in Escherichia coli prokaryotic expression system. Methods SjCL1 gene was amplified by PCR from plasmid pcDNA3 SjCL1 and cloned into prokaryotic expression vector pGEX 4T 1 to construct recombinant plasmid pGEX SjCL1. The recombinant plasmid was transformed into Escherichia coli JM109. Transformants were transfected with isopropyl β D thiogalactoside. The expression products of SjCL1 gene were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. As a result, a PCR fragment of about 1 kb was obtained and a pGEX SjCL1 plasmid was constructed. The expressed product was detected by SDS PAGE and Western blotting. The relative molecular mass was 6200. Conclusion SjCL1 gene is expressed in fusion form in Escherichia coli