【摘要】 目的研究多房棘球绦虫EmⅡ/3外源基因在大肠杆菌和BCG中的表达。方法将重组质粒pBCG-EmⅡ/3转入感受态大肠杆菌Top10和用电穿孔法导入BCG构建rBCG,将含有重组子的细菌培养至对数生长期,然后45℃热诱导,分别对表达产物作SDS-PAGE及免疫印迹分析。结果多房棘球绦虫pBCG-EmⅡ/3在BCG中成功表达了EmⅡ/3重组蛋白,在相对分子量为65 kDa处可见明显的表达蛋白带,其表达量占菌体总蛋白量的9%,且能与鼠血清抗体发生特异结合,而在大肠杆菌中没有外源蛋白表达。结论多房棘球绦虫pBCG-EmⅡ/3能在BCG中表达,Western blot结果提示rBCG-EmⅡ/3表达的EmⅡ/3重组蛋白具有特异的抗原性。更多还原

【Abstract】 To investigate the expression of gene encoding the EM II/3 antigen of Echinococcus multilocularis in E.coli and BCG,the recombinant plasmid pBCG-EM II/3 was transformed into the susceptible strain of E.coli Top10,and introduced to BCG by electroporation to construct the recombinant BCG.The bacteria containing the recombinants were cultivated to the logarithm growth period and induced by heating at 45℃.Meanwhile,the expressed products were analyzed by SDS-PAGE and Western blotting respectively.The experimental results showed that the plasmid pBCG-Em II/3 could successfully express the EM II/3 protein in BCG.which was identified as a band of 65 kDa as demonstrated by SDS-PAGE analysis,and the content of EM II/3 protein accounted for 9% of the total bacterial protein of BCG,This protein could bind with specific antibodies in mouse sera.However,the same kind of protein was not expressed in E.coli.It is concluded that the recombinant protein EM II/3 can be expressed in BCG and this kind of protein shows specific antigenicity.更多还原