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基于HIV-1pNL4-3构建含CRF07_BCV3的嵌合分子克隆

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Keywords: 人类免疫缺陷病毒,CRF07_BC,V3,嵌合病毒

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Abstract:

目的在感染性克隆pNL4-3骨架上构建含CRF07_BCV3的嵌合分子克隆。方法通过融合PCR将CRF07_BCpXJDC13的V3环与pNL4-3的env骨架进行融合并克隆至pNL4-3上。经过鉴定后将阳性V3嵌合克隆pNL4-3/XJDC13V3转染到293T细胞进行病毒包装并检测病毒的感染性。V3嵌合病毒的细胞嗜性分别用Ghost细胞系和MT-2细胞进行测定。结果利用融合PCR克隆方法在pNL4-3骨架上成功构建了含CRF07_BCpXJDC13V3的嵌合分子克隆pNL4-3/XJDC13V3。该嵌合病毒利用的辅助受体为CCR5,不能利用CXCR4,也不能在MT-2细胞上诱发合胞体。结论在pNL4-3骨架上成功构建了含CRF07_BCpXJDC13V3嵌合分子克隆pNL4-3/XJDC13V3。该嵌合分子克隆为进一步研究CRF07_BC嗜性转变中V3关键位点提供了有利工具。

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