EPEC O2、O78及融合双价弱毒菌株O2．78（Chl^r Nor^r）ompA基因原核表达载体重组构建

目的构建PThioHisA-ompA重组质粒。方法根据GenBank中大肠埃希菌K-12外膜蛋白A基因的核苷酸序列设计引物,从大肠埃希菌O2、O78及其融合株O2,78(ChlrNorr)中分别扩增得到外膜蛋白A基因(ompA)序列,进行序列测定及分析。重组PMD-18-ompA经双酶切切下ompA片段,电泳回收;原核表达载体PThioHisA质粒双酶切,回收大片段。再将回收的ompA插入回收大片段原核表达载体PThioHisA上。结果按预期PThioHisA-ompA重组质粒PCR产物经1%琼脂糖凝胶电泳,在约1kb处有一清晰条带,与预期值相符。结论成功构建PThioHisA-om-pA重组质粒,为致病性大肠埃希菌基因疫苗的抗原筛选和构建奠定了实验基础。

Construction of recombinant expression vector of ompA gene from EPEC O2, O78 and integration bivalent attenuated strain O2,78 (Chlr Norr)

Objective To construct the recombinant plasmid PThioHisA-ompA. Methods According to the nucleotide sequences of Escherichia coli K-12 ompA gene in GenBank, the primers were designed, and ompA from the E. coli O2,O78 and their integration of O2,78 (ChlrNorr ) were amplified, sequenced and analyzed. The recombinant plasmid pMD-18-ompA was digested and ompA was recoveried, and subcloned into prokaryotic expression vector PThioHisA plasmid. Results The ompA gene was successfully cloned into PThioHisA. Conclusion The constructed recombinant plasmid PThioHisA-ompA provids the basis for E. coli DNA vaccine antigen screening.