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目的:构建含CC细胞内趋化因子(CC-intrakine,RANTES-K)基因的慢病毒载体,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PCR技术,扩增目的基因片段RANTES-K,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-R-K,并在293FT细胞中建立慢病毒株,最后转染HeLa细胞观察RANTES蛋白的表达.结果:成功构建了慢病毒载体pLenti6/V5-R-K,并证实RANTES蛋白可在人宫颈癌HeLa细胞系内表达.结论:慢病毒载体可介导CC-intrakine(RANTES-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 containing CC-intrakine (RANTES-K) gene and lay a foundation for the study of gene therapy for HIV-1 infection.Methods: , And the target gene fragment RANTES-K was amplified. The purified PCR product was ligated into pLenti6 / V5-D-TOPO vector to construct lentiviral vector pLenti6 / V5-RK, and a lentivirus strain was established in 293FT cells. The expression of RANTES protein was observed by HeLa cells.Results: The lentiviral vector pLenti6 / V5-RK was successfully constructed and confirmed that RANTES protein was expressed in human cervical cancer HeLa cell line.Conclusion: Lentiviral vector can mediate the expression of CC-intrakine RANTES-K) gene is efficiently and stably transfected into HeLa cells for gene therapy of anti-HIV-1 infection.