about the people ccr5delta32 gene cloning and recombinant lentiviral vector construction（关于人民ccr5delta32基因克隆和重组慢病毒载体建设）.docVIP

About the people CCR5Delta32 gene cloning and recombinant lentiviral vector construction Papers network: Author: Ann Stars FORCES, Zhangxian Qing, Chen Rui, Zhang Ying, YU Rui Liu Ning [Abstract] Objective: human cloning CCR5Delta32 gene and gene therapy construct recombinant lentiviral vector containing the gene pLenti CCR5Delta32, for acquired immune deficiency syndrome (AIDS). Methods: from CCR5Delta32 mutations in individual peripheral blood mononuclear cells ( PBMCs within extraction of human genomic DNA using PCR technology amplified CCR5Delta32 full-length gene, after EcoRI single digestion with connected to the pUCm T carrier, followed by transformation of competent E. coli DH5 @, the extracted plasmid restriction analysis and DNA sequencing. again identified CCR5Delta32 subcloned into lentiviral vector pLenti6/V5 D TOPO and restriction enzyme digestion and DNA sequence analysis. Results: about 650 bp was obtained by PCR DNA fragment sequencing results with the GenBank sequence exactly cloned target gene has been inserted correctly into the lentiviral vector pLenti6/V5 D TOPO Conclusion: people CCR5Delta32 gene was successfully cloned and constructed recombinant lentiviral vector containing the gene pLenti CCR5Delta32 lay the foundation for further AIDS gene therapy research. [Keywords] people CCR5Delta32 gene, gene cloning, lentiviral vectors, HIV infection, AIDS [Abstract] AIM: To clone human CCR5Delta32 gene and construct the recombinant lentiviral vector pLenti CCR5Delta32 for the further gene therapy of AIDS. METHODS: Full length CCR5Delta32 gene was amplified by using PCR from human peripheral blood mononuclear cells (PBMCs) genomic DNA, and then cloned into pUCm T. After sequence analysis, the CCR5Delta32 gene was subcloned into lentiviral vector pLenti6/V5 D TOPO. RESULTS: A DNA fragment of 650 bp was obtained by PCR and its sequence was in conformity with the sequence reported in GenBank. The cloned gene was inserted into the lentivira