【摘要】 目的 :构建高免疫原性的抑制素真核表达载体。方法 :通过PCR扩增pCMV S基因中的S基因片段 ,然后将抑制素α(1 32 )片段与S的融合基因克隆到真核表达质粒pcDNA13.1( ) :酶切、测序鉴定后 ,采用脂质体包裹法将重组质粒转染Hela细胞 ,用SDS PAGE和ELISA对其表达产物进行检测。用抑制素融合表达质粒免疫大白鼠 ,ELISA检测血中抗抑制素抗体水平。结果 :酶切鉴定和序列分析表明 ,融合基因的表达质粒pClS构建成功。表达质粒在HeLa细胞中获得了表达 ,融合蛋白的分子量约为 2 9kD ,融合蛋白具有抑制素免疫原性。重组质粒免疫使大白鼠产生了抗抑制素抗体。结论 :高免疫原性的抑制素表达质粒的构建为利用抑制素基因免疫技术诱导单胎动物生多胎奠定了基础。更多还原

【Abstract】 Objective:To construct inhibin expression vector with high immunogenicity.Methods:S gene with enzyme site sequence of EcoR I and Nde I was amplified by PCR from pCMV-S;Inhibin(1-32)gene with enzyme site sequence of Nde I and Hind III was cut from pUI;S and inhibin(1-32)gene were directly inserted into pcDNA3.1(-)and the recombinant pCIS was identified by restriction endonculease digestion and sequencing;then pCIS was transfected into HeLa cells.SDS-PAGE and ELISA was employed to detect the cell product.Rats were immunized with inhibin fused expression plasmid pCIS and ELISA was used to detect the blood antibody against inhibin.Results:The recombinant eukaryocyte expression vector of inhibin was constructed.The sequence of the insert was identical to designed gene.Fusion protein weight was about 29 kD and it had inhibin immunologic activity.Rats immunized with pCIS induced antibody against inhibin.Conclusion:The fusion gene expression vector was successfully constructed,and it set up the basis of inhibin gene immunization to induce multiple bear for single birth animals.更多还原