【摘 要】
：
目的:构建腺病毒纤维蛋白基因的真核表达质粒,并检测其在真核细胞中的表达,为腺病毒靶向性载体构建创造条件.方法:采用限制性内切酶技术,酶切腺病毒骨架质粒pAdEasy-1,经多次
【机 构】
：
汕头大学医学院生化与分子生物学教研室,广东,汕头,515031第四军医大学全军消化病研究所;西安交通大学第二医院耳鼻喉科,陕西,西安,710032;
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目的:构建腺病毒纤维蛋白基因的真核表达质粒,并检测其在真核细胞中的表达,为腺病毒靶向性载体构建创造条件.方法:采用限制性内切酶技术,酶切腺病毒骨架质粒pAdEasy-1,经多次亚克隆,最后完成克隆纤维蛋白基因并构建其真核表达质粒.用脂质体法将构建好的真核表达质粒瞬时转染COS-7细胞,于转染后72 h,用Western blot法检测所表达的蛋白.结果:克隆成功纤维蛋白基因,并构建纤维蛋白基因真核表达质粒pcDNA/Fiber,经限制性内切酶酶切鉴定及测序证实了其正确性.Western blot结果显示,新表达蛋白在变性条件下大小为62 kD,而在非变性条件下为186 kD.结论:纤维蛋白基因真核表达质粒能在真核细胞中表达,产物具有三聚体结构,可用于腺病毒靶向性载体的构建.
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