变形链球菌表面蛋白可变区(V+)基因克隆与表达

Cloning and Expression of Variable Domain (V+) Gene of Surface Protein of Streptococcus Mutans

摘要: 目的分离变形链球菌表面蛋白可变区(V+)基因,构建V+表达克隆.方法用PCR从sr基因中扩增目的基因片段V(+),亚克隆入中间载体pCR2.1和表达载体pET21a(+).对重组体进行诱导表达并对表达产物进行SDS-PAGE和Western blotting分析.结果PCR扩增产物为116kb的DNA带,亚克隆入载体,获得重组克隆pCVf和重组表达克隆pSRVf,表达产物为43kDa抗原,可以被抗Ⅰ/Ⅱ抗体特异性地识别.结论本试验成功地构建了携带V+基因片段的表达克隆pSRVf.

Abstract: ObjectiveTo isolate variable domain(V+)gene of the surface protein of streptococcus mutans and construct an expressive clone of V+gene.MethodsV+do main was amplified from gene sr by poly merase chain reaction(PCR)and subcloned into the plasmid vector pCR2.1 and the expressive plasmid vector pET21a(+).The recombinant was induced by IPTG to express target protein which was analysed by SDS-PAGE Western blotting.ResultsThe specific product of PCR was about 1.16bp which was subcloned into vectors.The recombinant clone pCVf and the recombinant expressive clone pSRVf were obtained.pSRVf expressed specific protein with a molecular weight of 43kDa.This recombinant protein could be specifically recog nized by anti Ⅰ/Ⅱ serum.ConclusionV+domain of sr gene was cloned successfully.