旋毛虫Ts87基因在大肠杆菌中的表达及表达产物的纯化

Expression and purification of Ts87 gene of Trichinella spiralis

目的 为进一步研究和应用旋毛虫Ts87基因 ,本文原核表达了PET - 2 8a(+) /Ts87重组质粒 ,并纯化鉴定了该蛋白。方法 将Ts87基因片段克隆入原核表达载体PET - 2 8a(+) ,形成重组质粒PET - 2 8a(+) /Ts87,转化后经IPTG诱导得到表达蛋白 (约 4 0kDa) ,利用固定化金属离子 (Ni2 + )配体亲和层析纯化目的蛋白 ,进行复性 ,并用兔人工感染旋毛虫血清鉴定表达产物。结果与结论 成功构建了PET - 2 8a(+) /Ts87,SDS -PAGE显示融合蛋白主要以包涵体形式存在 ,经过亲和层析得到纯化的目的蛋白 。

Aim To establish a expression purification procedure for Ts87 gene in vitro Methods After centrifugation of the bacterial culture,the bacterial pellet was resuspended and lysed by ultrasonication Immobilized metal (Ni 2+ ) chelation affinity chromatography was used to purify the fusion protein PET-28a(+)/Ts87 from the inclusion bodies Results and Conclusion The SDS-PAGE analysis showed that the fusion protein was stayed as inclusion bodies The target protein was purified by affinity chromatography The purified fusion protein could react with Trichinella spiralis infected rabbit sera by Western blot