B型肉毒毒素受体结合区C片段在大肠杆菌中的表达及免疫原性研究

Expression in Escherichia coli and Immunogenicity of C Fragment of Receptor Binding Domain of Botulinum Nurotoxin Serotype B

目的:在大肠杆菌中表达、纯化B型肉毒毒素受体结合区C片段(BHc-C),研究其免疫原性。方法:将BHc-C基因克隆到原核表达载体pGEX-4T-1中,转化大肠杆菌BL21(DE3),经IPTG诱导表达GST-BHc-C融合蛋白并通过亲和纯化;以纯化的融合蛋白免疫BALB/c小鼠制备免疫血清,采用ELISA检测免疫血清的效价并测定其抗B型肉毒毒素中和活性。结果:在大肠杆菌中表达了GST-BHc-C融合蛋白;以该融合蛋白免疫小鼠获得高效价免疫血清,且该免疫血清具有中和活性。结论:获得了GST-BHc-C融合蛋白,并证实其具有免疫原性。

Objective：To express recombinant C fragment of receptor binding domain（BHc-C） of botulinum nuro-toxin serotype B（BoNT / B） in Escherichia coli and explore its immunogenicity.Methods：The gene encoding C fragment of receptor binding domain of BoNT / B was cloned into prokaryotic GST fusion expression vector pGEX-4T-1.The recombinant plasmid was introduced into E.coli BL21（DE3） and fusion protein GST-BHc-C was ex-pressed and purified.Then,the purified GST-BHc-C was used to vaccinate BALB / c mice,serum antibody was detected by EILSA and protective antibody was also determined by the BoNT / B neutralization assay.Results：The fusion protein GST-BHc-C was successfully expressed in E.coli.BALB / c mice vaccinated with GST-BHc-C in-duced a strong and specific antibody.The serum from vaccinated mice contained neutralization antibody against BoNT / B.Conclusion：The fusion protein GST-BHc-C with strong immunogenicity is obtained through the GST fu-sion expression system.