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目的:构建人S100A8基因高表达的重组慢病毒表达载体。方法:以正常人骨髓单个核细胞的c DNA为扩增模板,克隆S100A8基因全长的c DNA,与p MD19-T载体连接后进行测序鉴定;将p LVX-IRES-puro目的载体与测序正确的S100A8片段进行连接,筛选阳性克隆后进行菌落PCR、双酶切及测序鉴定。结果:p LVXS100A8重组质粒双酶切及菌落PCR产物与目的基因相关片段S100A8 c DNA大小一致,成功克隆S100A8基因,S100A8基因成功插入到p LVX-IRES-puro。结论:成功构建了人S100A8基因高表达的重组慢病毒表达载体。
Objective: To construct a recombinant lentiviral vector with high expression of human S100A8 gene. Methods: The c DNA of S100A8 gene was cloned from c DNA of normal human bone marrow mononuclear cells and cloned into pMD19-T vector and sequenced. The vector p LVX-IRES-puro was sequenced correctly Of S100A8 fragments were connected, after screening positive clones colony PCR, double enzyme digestion and sequencing identification. Results: The size of S100A8 cDNA of pVXX100A8 recombinant plasmid was identical with that of S100A8 cDNA. The S100A8 gene was successfully cloned and S100A8 gene was successfully inserted into pVXX-IRES-puro. Conclusion: The recombinant lentiviral vector with high expression of human S100A8 gene was successfully constructed.