猪带绦虫六钩蚴免疫原基因的克隆、表达与结构预测

The Cloning & Expression of Immunogen Gene of Oncosphere of Taenia solium and Structural Prediction of Protein

从猪带绦虫六钩蚴cDNA表达文库中筛选、克隆的Ts01基因,设计表达性引物,亚克隆到pGEX-4T-1表达载体,构建了pGEX-GST-Ts01的融合表达重组载体,转化BL21寄主菌和IPTG诱导表达后,经SDS-PAGE和Western蛋白分析,表明成功地高效表达了约40000左右的融合蛋白质,与预测的相对分子质量大小一致,表达量达40%左右。表达产物具有免疫学活性,而且呈可溶性,未形成包涵体,这与用蛋白质软件分析的Ts01编码蛋白水溶性相一致。

The interesting gene of Ts01 from library was cloned to expression vector of pGEX-4T-1 with special primer,and constructed the fusion expression vector of pGEX-GST-Ts01 and then transformed into E.coli of BL21.The fusion expression of Ts01 was induced with IPTG,the expression product was analyzed by SDS-PAGE and Western-blot.The results showed that the Ts01 fusion protein has been expressed successfully in BL21.The fusion protein was about 40 000 in Mr and reached 40% of expression level.It can be distinguished by positive serum of cysticercosis.The product was a soluble molecule and accord to those predicted by protein analysis soft.