牛分枝杆菌Mb1761c蛋白的表达及其免疫原性初步研究

Expression of Mb1761c of M.bovis and initial research on its antigenicity

根据GenBank提供的M.bovis Mb1761c基因序列设计引物。以牛分枝杆菌DNA为模板,通过PCR方法扩增出牛分枝杆菌MB1761c基因片段,以BamHⅠ和EcoRⅠ双酶切PCR产物和pET28a(+)后将纯化的Mb1761c基因克隆至pET28a(+)中,构建出原核表达质粒,再转化到E.coli BL21(DE3)表达菌株中,IPTG诱导后,收集菌体进行SDS-PAGE,分析可见约29ku特异性蛋白条带,Western blot分析发现,该融合蛋白具有较强的免疫原性,从而为进一步研究其亚单位疫苗及DNA疫苗奠定了基础。

The MB1761c gene of M.bovis was amplified from M.bovis genomic DNA by PCR and cloned into pET-28a （＋）. The recombinant plasmid was transformed into E.coli BL21 （DE3） and protein expression was induced by IPTG. Cell lysates were analyzed by SDS-PAGE and Western blot assay. The results showed that the expressed MB1761C was about 29 ku and possessed the antigenicity of the MB1761c protein. This work could serve as a basis for further studies on subunit vaccine and DNA vaccine against M. bovis.