副结核分枝杆菌hsp65基因在大肠杆菌中的表达

Expression of Mycobacterium Paratuberculosis hsp65 gene in Escherichia coli

本研究以EcoR Ⅴ和EcoR Ⅰ双酶切pGEM-T-hsp65和pET-32a(+),并将纯化的hsp65基因亚克隆至pET-32a(+)中,构建出原核表达载体pET-hsp65。将pET-hsp65转化至感受态E.coli BL21(DE3)中,经IPTG诱导和SDS-PAGE分析,可见一条约76ku的外源蛋白带。Western blot分析表明,该蛋白具有与副结核分枝杆菌多克隆抗体发生反应的特性,为进一步研究有关hsp65的新型疫苗奠定了基础。

The hsp65 gene of Mycobacterium avium subsp. Paratuberculosis （MAP） was cloned into the prokaryotic expression vector pET-32a-and transformed into the competence E.coli BL21 cells （DE3）. Expression of hsp65 gene was induced by IPTG and analyzed by SDS-PAGE. The expressed protein was about 76 ku and could detect by Western blot using MAP specific antibody. These results could serve as a basis for further studies on the MAP hsp65 gene and its development of new vaccines.