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登革Ⅱ型病毒Tr1751株E蛋白基因的克隆与表达载体的构建

, PP. 1050-1054

Keywords: 登革Ⅱ,病毒,E蛋白,克隆,真核质粒

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Abstract:

目的从登革Ⅱ病毒Tr1751株中克隆表达E蛋白的基因,构建了真核表达质粒,为其后继的关于结构和功能的研究提供了条件。方法合成DEN2(Tr1751株)的E蛋白基因引物,通过RT-PCR的方法扩增含有信号肽E蛋白的基因片段,并与真核载体pIRES-hrGFP-1α连接,得到重组的质粒pIRES-hrGFP-1α-E,再用PCR、双酶切和测序的方法进行鉴定。结果RT-PCR得到的为1527bp的E蛋白基因片段,酶切鉴定和测序显示重组的质粒含有DEN2病毒中的E蛋白的基因。用重组质粒测序后与GenBank中的已知Tr1751株进行相似性比对,核苷酸序列和推导的氨基酸序列同源性均为99%。结论成功的构建出含有全长DEN2的E蛋白基因的真核质粒,为登革病毒核酸疫苗的研究奠定了基础。

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