日本血吸虫p50亲免素基因克隆、表达及纯化  

Clone and expression of schistosoma japonicum p50 immunophilin gene and purification of p50 protein

作　　者：李孜[1] 余新炳[1] 吴忠道[1] 徐劲[1] 胡旭初[1] 

机构地区：[1]中山大学基础医学院寄生虫学教研室

出　　处：《中国公共卫生》2005年第11期1303-1305,共3页Chinese Journal of Public Health

基　　金：国家教育部博士点基金(200045);广东"211工程"重点建设项目(98169)

摘　　要：目的扩增、原核克隆和表达日本血吸虫(Sj)p50亲免素基因全长cDNA,并进行表达产物的纯化。方法从成虫RNA中RT-PCR扩增Sjp50亲免素基因完整的编码阅读框后,将其克隆入pET 30 a(+)原核表达载体中,异丙基硫代-β-D半乳糖苷(IPTG)诱导高表达后用亲和层析柱对表达产物进行纯化。结果将Sjp50亲免素基因成功克隆入pET 30 a(+)表达载体中;诱导表达并成功纯化出重组Sjp50亲免素蛋白。结论成功克隆Sjp50亲免素基因,并表达和纯化得到了p50亲免素蛋白,为研究Sjp50亲免素的功能及其进行疫苗等研究奠定了基础。Objective To clone and express Schistosoma japonicum （Sj） p 50 immunophilin gene, and to purify its protein. Methods To amplify p 50 immunophilin＇s complete coding sequence（CDS） from adult worm mRNA by RT-PCR. cDNA of p 50 immunophilin gene was cloned into the pET 30 a（＋）vector. The recombinant plasmid was transformed into E.coli BL 21 （DE 3） and expressed, then the expressed products in E.coli were purified by affinity chromatography column. Resuits p 50 immunophilin gene cDNA sequence was obtained and then successfully cloned into pET 30 a（＋）vector. p 50 immunophilin fusion protein was expressed after induced by IPTG and purified with （His）6 at the N terminus. Conclusion Sjp 50 immunophilin gene was successfully cloned, expressed and fusion protein was purified, which gave the basis for further study especially as a vaccine.

关 键 词：日本血吸虫 中国大陆株 p50亲免素 克隆 表达纯化 

分 类 号：R184.38[医药卫生—流行病学]