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目的构建含趋化因子融合蛋白SDF-KDEL的慢病毒载体pLenti6/V5-S-K,为研究I型人免疫缺陷病毒(HIV-1)感染的基因治疗奠定基础。方法应用聚合酶链反应(PCR)技术扩增目的基因片段SDF-KDEL,纯化后的PCR产物定向连接入pLenti6/V5-D-TOPO(载体,构建慢病毒载体pLenti6/V5-S-K,并在293FT细胞中建立慢病毒株,最后转染人宫颈癌HeLa细胞观察SDF-1蛋白的表达。结果成功构建了慢病毒载体pLenti6/V5-S-K,并证实SDF-1蛋白可在HeLa细胞系内表达。结论慢病毒载体可介导CXC-细胞内趋化因子(CXC-in-trakine,SDF-K)基因高效、稳定转染HeLa细胞,可用于抗HIV-1感染的基因治疗。
Objective To construct a lentiviral vector pLenti6 / V5-S-K containing the chemokine fusion protein SDF-KDEL, which lays a foundation for the study of gene therapy for HIV-1 infection. Methods The gene fragment SDF-KDEL was amplified by polymerase chain reaction (PCR). The purified PCR product was ligated into pLenti6 / V5-D-TOPO vector to construct lentiviral vector pLenti6 / V5- The lentiviral vector was constructed and transfected into HeLa cells to observe the expression of SDF-1 protein.Results The lentiviral vector pLenti6 / V5-SK was successfully constructed and confirmed that SDF-1 protein was expressed in HeLa cell line. CONCLUSION: Lentivirus vector can mediate efficient and stable transfection of CXC-in-trakine (SDF-K) gene into HeLa cells for gene therapy of HIV-1 infection.