【摘要】 目的构建艰难梭菌毒素A和毒素B的原核表达载体,获得融合表达抗原,并鉴定其抗原活性。方法以ATCC43255菌株基因组DNA为模板,通过PCR扩增获得毒素A和B羧基端(C端)基因片段,并进行原核表达获得毒素A和B羧基端抗原区段。利用艰难梭菌毒素检测试剂盒对毒素A和B羧基端抗原区段的抗原性进行鉴定。结果成功构建了艰难梭菌毒素A和毒素B的原核表达载体,并获得能被相应特异性抗体所识别的毒素A和B羧基端抗原区段。结论获得毒素A和B羧基端抗原区段具有很好的抗原活性,为进一步制备相应抗体并建立艰难梭菌毒素A和B的免疫检测方法奠定了基础。更多还原

【Abstract】 Objective To construct the prokaryotic expression vector of carboxyl( C)-terminal domains of Clostridium difficile toxins A and B,express recombinant antigens in Escherichia coli, and identify their antigenicity. Methods The genes of C-terminal domains of C. difficile toxins A and B were cloned from ATCC43255 genome DNA. The recombinant antigens were expressed in E. coli with IPTG induction and purified by Ni-NAT deads. The antigenicity was detected using C. diff Qick Chek Complete dual-antigen EIA. Results Prokaryotic expression vectors of C-terminal domains of C. difficile toxins A and B were constructed. The obtained C-terminal antigens could be identified by specific anti-toxins A and B antibodies. Conclusion The obtained C-terminal antigens of C. difficile toxins A and B can be used to prepare corresponding antibodies,which will help develop the immunoassay for C. difficile toxins A and B.更多还原