【摘要】 目的 :构建表达粘膜佐剂霍乱毒素B亚单位 (CTB)基因的重组质粒 ,并在大肠杆菌中表达获得基因重组蛋白。方法 :用PCR方法从霍乱弧菌中扩增CTB片段 ,将其转入原核载体质粒pET-32a( +) ,在大肠杆菌Top10克隆 ,并在BL21中表达。结果 :重组质粒PET-CTB的全长序列经分析与基因文库公布的一致 ;表达蛋白经SDS-PAGE分析 ,相对分子量与文献相符 ;重组蛋白竟Westernblot检测有抗原性。结论 :基因重组菌表达的CTB蛋白可能作为有效、安全的粘膜佐剂用于口服疫苗的研制更多还原

【Abstract】 Objective:To construct the recombinant plasmid of mucosa adjuvant cholera toxin subunit B (CTB) and express fusion protein in E.coli BL21.Methods:The gene of CTB was amplified from Vibrio cholerae.The gene was inserted into the prokaryotic expression vector pET-32a(+).The recombinant CTB was expressed in E.coli BL21.Results:The DNA sequence of recombinant pET-CTB was identified with the sequence published by GenBank.The relative molecular mass (Mr) of fusion protein was corresponding to Mr shown in SDS-PAGE.The immunogenicity of recombinant CTB was confirmed by western blot analysis.Conclusion:The recombinant protein CTB may function as oral vaccine effective and safe.更多还原