华支睾吸虫GST2-TP22.3融合蛋白的构建及其免疫原性初步研究

目的构建华支睾吸虫候选诊断分子pET32a（＋）-GST2-TP22.3重组质粒,表达及纯化CsGST2-CsTP22.3融合蛋白,为诊断抗原的研究奠定基础。方法选用合适的限制性内切酶,先后将CsGST2、CsTP22.3克隆入pET32a（＋）质粒,转化BL21宿主菌,IPTG诱导重组质粒的表达,亲和层析纯化表达产物,用SDS-PAGE和Western blotting进行分析鉴定。结果 PCR、双酶切、测序结果均表明CsGST2-CsTP22.3重组质粒构建成功,重组融合蛋白可被其免疫的SD大鼠血清识别,表明其具有免疫原性。结论 pET32a（＋）-CsGST2-CsTP22.3重组质粒构建成功,为下一步研究其功能奠定基础。

Preparation of fusion protein GST2-TP22.3 of Clonorchis Sinensis and analysis of immunogenicity of the recombinant protein

To construct and express CsGST2-CsTP22.3 fused protein of Clonorchis sinensis,and evaluate its applicability in serological diagnosis of clonorchiasis,CsGST2 and CsTP22.3 genes were successively cloned into the prokaryotic expression vector pET32a（＋） and then expressed in E.coli BL21 by IPTG induction.Then the recombinant CsGST2-CsTP22.3 fusion protein was expressed and identified by SDS-PAGE and Western Blotting.PCR,double enzyme digestion and DNA sequencing confirmed that the recombinant expression plasmid was successfully constructed.The recombinant fusion protein could elicit efficiently specific antibodies in SD rats,indicating its ability of immunogenicity and laying the foundation for further study of its function.