中文摘要：

为观器重组MPB83蛋白的免疫活性，揭示该蛋白在牛结核病的诊断和防治中的作用，克隆了牛分枝杆菌MPB83基因，构建了屯隆载体pGEMMPB83和枉达载体pET30a MPB83，经IPTG诱导在大肠杆菌BL21（DE3）中表达，用SDS-PAGE和免疫印迹分析表达产物并进行蛋白纯化。试验结果丧明：牛分枝杆菌MPB83基因体外扩增产物与预期值相符。约600bp；所构建表达质粒pET30a。MPB83经测序，结果与预期一致；SDS-PAGE分析表明，该融合蛋白以包涵体的形式表达．其分子质量约为26ku，蛋白缸达量占菌体总蛋白的20％；该蛋白经电洗脱纯化后，纯度达95％以上；免疫印迹分析表明．原核表达的融合蛋白可与兔抗牛分枝杆菌多克隆抗体结合，并且具特异的免疫反应性。

英文摘要：

In order to determine the function of the MPB83 in the prevention and diagnosis of tuberculosis in cattle, the MPB83 gene of Mycobacterium boris was cloned. Then vector pGEM-MPB83 and prokaryotic expression vector pET30a-MPB83 were constructed. Recombinant E. coil BL2I （DE3） was induced by IPTG to express the fusion protein. The expressed and purified product was analyzed by SDS-PAGE and Western-Blot. The result showed that PCR product was about 600 bp as expected. Expression vector pET30a-MPB83 was conformed confirmed by sequencing. The results of SDS-PAGE showed that the fusion protein was produced abundantly as inclusion body and the molecular weight of expressed protein was about 26 ku. SDS-PAGE analysis also showed that the recombinant protein expressed could reach 20 percent of the whole bacterial protein. Purified protein was obtained after being eluted from the gel by electrophoresis. The Western-blotting analysis showed the fusion protein had the antigenic activity of Mycobacterium boris.