日本血吸虫SJCHGC02377蛋白部分片段的克隆与表达

目的:克隆和表达日本血吸虫SJCHGC02377蛋白。方法:采用RT-PCR从日本血吸虫成虫总RNA中扩增出编码SJCHGC02377蛋白的基因片段;经克隆和测序分析后,亚克隆至表达载体pET-23 a(+),转化大肠埃希菌BL21,菌落PCR和双酶切鉴定;阳性菌株经IPTG诱导表达SJCHGC02377蛋白部分片段,亲和层析予以纯化。结果:RT-PCR扩增出SJCH-GC02377蛋白基因片段;获得的阳性克隆序列与GenBank中登录的日本血吸虫SJCHGC02377蛋白的基因序列一致;SJCH-GC02377蛋白基因被亚克隆到原核表达载体pET-23 a(+),在BL21中获得了表达,表达的蛋白经亲和层析获得纯化。结论:成功构建了日本血吸虫SJCHGC02377蛋白的重组表达质粒,在大肠埃希菌中获得表达。

Cloning and Expression of Schistosoma Japonicum Truncated SJCHGC02377 Protein

Objective To clone and express Schistosoma japonicum SJCHGC02377 protein.Methods The gene encoding Schistosoma japonicum SJCHGC02377 protein was amplified by RT-PCR from Schistosoma japonicum worm total RNA,and was cloned into pMD18-T vector.Positive clones were identified by colony PCR and sequencing with ABI PRISMTM 377XL DNA Sequencer;The insert digested with BamHⅠand SalⅠwas subcloned into prokaryotic expression vector pET-23a(+) which were also digested with BamHⅠand SalⅠ;The recombinant plasmids were ...