利用昆虫细胞杆状病毒表达系统表达重组人类ZP3蛋白

来源 :GTAUF2014 & APSMHA2014第六届长城泌尿男科转化医学论坛暨第七届亚太男性健康与抗衰老研究学会年会 | 被引量 : 0次 | 上传用户：zy197855

【摘要】

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【作者】

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卢慧刁华肖玉芳于合国李铮施惠娟

【机构】

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上海交通大学医学院附属仁济医院泌尿外科,上海市人类精子库,上海200135,中国上海市计划生育科

【出处】

：

GTAUF2014 & APSMHA2014第六届长城泌尿男科转化医学论坛暨第七届亚太男性健康与抗衰老研究学会年会

【发表日期】

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2014年11期

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目的:用人类ZP3来评估精子ZP结合能力是一个有效手段,但人类ZP3蛋白来源非常有限,限制了ZP3蛋白在精子质量评估和辅助生殖中的应用.探讨利用重组表达系统表达制备重组人类hZP3蛋白的方法和技术难点,为hZP3蛋白的制备和应用打下技术基础.材料方法:构建重组人hZP3的载体质粒pFASTBAC HTa-hZP3,在大肠杆菌宿主细胞中利用杆状病毒Bac-to-Bac系统同源重组构建重组的杆状病毒穿梭载体Bacmid,用重组的Bacmid转染sf9细胞制备杆状重组病毒,利用重组的杆状病毒感染sf9细胞,分别表达带HIS标签的人类ZP3全长蛋白(氨基酸1-424)和截短的hZP3蛋白(氨基酸23-348).

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