【摘要】 目的为建立人巨细胞病毒(HCMV)活动性感染检测方法,表达HCMV早期复制的标志物pp65蛋白,以制备相应的抗体。方法将pp65全基因和其亲水性强、有抗原代表性的片段(pp65hp)分别克隆到表达载体pET22b(+)中诱导表达,并以免疫印迹考察重组蛋白的抗原性。结果重组rpp65全蛋白表达量低于全菌体蛋白的5%,而rp65hp蛋白获得高效表达,约为全菌体蛋白的40%,并与HCMVIgG抗体特异性结合。结论亲水性pp65hp片段有较好的抗原性,可以制备相应的抗体,来检测HCMV活动性感染。更多还原

【Abstract】 Objective To detect the active infection of HCMV .The early infection marker pp65 was expressed and used for preparing antibody.Methods The expression vectors containing whole pp65 gene and one hydrophilic and antigenic domain of pp65(pp65 hp) were constructed and induced to express in E.coli, respectively. Western Bolt was applied to test the antigenicity of recombinant protein.ResultWhole pp65 was expressed at low level, less than 5% of total bacteria protein; while high efficient expression was observed for rp65 hp, approaching 40% of total protein in bacteria. Western Bolt result showed that rp65hp reacted with HCMV IgG antibody.Conclusion Recombinant rp65hp was antigenic and can be used to prepare antibody for detection of early HCMV infection.更多还原