【摘要】 为构建高免疫原性的卵泡抑制素(Inhibin,INH)DNA疫苗,将INH基因片段α1-32插入到pcIS中乙肝表面抗原(HBsAg)S基因第112~113氨基酸残基密码子之间,构建含2拷贝INH的融合表达质粒pcISI。酶切和测序鉴定表明,重组质粒pcISI构建成功。脂质体包裹法将pcISI转染HeLa细胞,ELISA检测表达产物的INH免疫反应原性。结果表明,融合目的基因在HeLa细胞获得表达,ISI融合蛋白具有比IS融合蛋白更强的INH抗原抗体反应性。将pcISI免疫6只大鼠后,5/6的大鼠产生了抗INH抗体,抗体P/N值在免疫后第2~6周高于pcIS免疫组。这些结果表明,所构建的质粒pcISI可以表达抑制素,表达产物具有较强的免疫原性。更多还原

【Abstract】 To construct an inhinbin(INH) expression vector with high immunogenicity,INHα1-32 gene was incorporated into the S gene of pcIS between amino acid residues 112 and 113.The resultant plasmid pcISI with two copies of INHα1-32 was identified by restriction endonuclease digestion and sequencing.The plasmid pcISI was then introduced into HeLa cells via liposome-mediated transfection and the immunogenicity of recombinant protein was detected by ELISA.The results showed that the recombinant gene expressed in HeLa cells,and the INH immunogenicity of ISI fusion protein was higher than that of IS fusion protein,indicating the ISI expression vector was constructed successfully.Five of six rats immunized with pcISI produced antibodies against INH in vivo.In addition,the P/N values of antibodies were higher than those immunized with pcIS from 2nd week to 6th week.In conclusion,the constructed pcISI could express INH with high immunogenicity.更多还原