日本血吸虫BBC1基因的克隆、表达及鉴定  

CLONING, EXPRESSING AND IDENTIFICATION OF BBC1 GENE OF SCHISTOSOMA JAPONICUM

作　　者：万志刚[1] 肖建华[1] 曾桥[1] 张愉快[1] 

机构地区：[1]南华大学病原生物研究所,湖南衡阳421001

出　　处：《中国寄生虫病防治杂志》2005年第4期282-284,共3页Chinese Journal of Parasitic Disease Control

摘　　要：目的克隆和表达日本血吸虫BBC1(SjBBC1)基因,为寻找新的抗日本血吸虫感染候选抗原分子提供实验基础。方法用PRIMER5.0引物设计软件自行设计引物,PCR扩增SjBBC1基因,将PCR产物纯化后与pUCm-T载体连接,经蓝白筛选、双酶切分析和PCR鉴定后,亚克隆入pQE30原核表达载体中。对原核表达产物进行SDS-PAGE和Western-blot鉴定。结果PCR扩增产物约为650bp,与预期大小相符。将该基因克隆入原核表达载体pQE30,表达蛋白分子质量单位约为23.6ku,并能被日本血吸虫感染兔血清识别。结论构建了pQE30/SjBBC1原核表达重组体载体,表达了具有抗原性的BBC1抗原。Objective To clone and express the Schistosornajaponicurn gene-BBC1（SjBBC1）, so as to look for new candidate antigen of resisting S. japonicum. Methods A pair of primers was synthesized after being designed by PRIMERS. 0 software according to the cDNA sequence of SjBBC1 gene. The SjBBC1 gene was amplified by PCR and was inserted into the cloning vector of pUCm-T. The fragment of SjBBC1 inserted into pUCm-T was subcloned into prokaryotic expressed vector pQE30. The recombinant plasmids of pQE30/SjBBC1 was screened by ampicilin and identified with restrictive enzymes. The SjBBC1 gene was transformed into E. coli DHSa and espressed with IPTG inducing. The expressed products were identified by SDS-PAGE and Western blot. Results The amplified fragment was 650 bp by agarose gel electrophoresis and sequencing. The fragment of SjBBC1 inserted into pUCm T was subcloned into prokaryotic expressed vector pQE30 and a peptide of about 23.6 ku was obtained. Conclusion The recombinant piasmid of pQE30/ SjBBC1 was constructed and the BBC1 protein was expressd in E. coll.

关 键 词：血吸虫 日本 BBC1基因 基因克隆 表达 

分 类 号：R383.24[医药卫生—医学寄生虫学]