目的在成功构建猪带绦虫大肠杆菌-双歧杆菌穿梭表达质粒pGEX-TSOL18的基础上,研究猪带绦虫TSOL18基因在长双歧杆菌中的表达情况。方法将猪带绦虫大肠杆菌-双歧杆菌穿梭表达质粒pGEX-TSOL18电转化入长双歧杆菌,IPTG诱导表达,SDS-PAGE和Western blot分析表达情况。结果酶切、PCR和测序证实,重组质粒pGEX-TSOL18成功转入长双歧杆菌。SDS-PAGE显示,目的蛋白相对分子质量(Mr)约为41KD,与预期结果相一致。Western blot显示,目的蛋白能被兔抗血清、囊虫病猪血清和囊虫病患者血清所识别。结论猪带绦虫TSOL18基因能够在长双歧杆菌中获得表达,表达的目的蛋白具有抗原性。
Objective On the basis of successful construction of the E.coil-B.longum shuttle plasmid pGEX-TSOL18 of Taenia solium,the expression of the gene of TSOL18 of Taenia solium was observed in B.longum.Methods The shuttle expression plasmid pGEX-TSOL18 of Taenia solium was electroporated into B.longum.After induction with IPTG,the expression of the recombinant protein was identified by SDS-PAGE and Western blot.Results Restriction analysis,PCR and sequencing showed that the recombinant plasmid pGEX-TSOL18was successfully transformed into B.longum.SDS-PAGE analysis showed that the relative molecular mass(Mr)of the target protein was approximately 41KD.The rabbit antiserum,cysticercosis swine serum and cysticercosis patients serum could bind to the target protein in Western blot assay,respectively.Conclusion The gene of TSOL18 of Taenia solium could be expressed in B.longum,and the expressed target protein showed antigenicity.