猪带绦虫TSO45W-4B-TSOL18融合基因在长双歧杆菌中的表达

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-TSO45W-4B-TSOL18的基础上,研究猪带绦虫TSO45W-4B-TSOL18融合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-TSO45W-4B-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-TSO45W-4B-TSOL18成功转入长双歧杆菌。SDS-PAGE显示,重组蛋白相对分子质量(M_r)约为55 kD,与预期结果相一致。Western blot显示,重组蛋白能被兔抗血清、囊虫病猪血清和囊虫病患者血清所识别。结论 猪带绦虫TSO45W-4B-TSOL18融合基因能够在长双歧杆菌中获得表达,表达的重组蛋白具有特异的抗原性。

Taenia solium TSO45W-4B-TSOL18 fusion gene expression in Bifidobacterium longum

On the basis of successful construction of the Escherichia coli （E. coli）-Bifidobacterium shuttle plasmid pGEX-TSO45W-4B-TSOL18 of Taenia solium, the expression of the fusion gene TSO45W-4B-TSOL18 of Taenia solium was observed in B. longum （Bifidobacterium longum）. The shuttle expression plasmid pGEX-TSO45W-4B-TSOL18 of Taenia so- lium was electroporated into B. longum. After induction with IPTG, the expression of the recombinant protein was identified by SDS-PAGE and Western blot. Restriction analysis, PCR and sequencing showed that the recombinant plasmid pGEX- TSO45W-4B-TSOL18 was successfully transformed into B. longum. As demonstrated by SDS-PAGE, the relative molecular mass （Mr） of the expressed recombinant protein was approximately 55 kD. The rabbit antiserum, cysticercosis swine serum and cysticercosis patients＇ serum could bind to the recombinant protein in Western blot assay. It was demonstrated that the fu- sion gene TSO45W-4B-TSOL18 of Taenia solium could be expressed in B. longurn, and the expressed recombinant protein shows specific antigenicity.