人S100A8基因重组慢病毒表达载体的构建

Construction of Recombinant Lentivirus Expressing Vectors of Human S100A8 Gene

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 the recombinant lentiviral expression vector of human S100 A8 gene high expression. Methods： The marrow mononuclear cells c DNA as template in the normal human bone,the full-length c DNA of cloned S100 A8 gene was connected to the p MD19-T vector and identified by sequencing. The p LVX-IRES-puro target vector was connected with the sequenced S100 A8 fragment. After screening the positive clones,colony PCR,double enzyme digestion and sequencing were performed. Results： PLVX-S100 A8 recombinant plasmid double enzyme digestion and colony PCR products have the same size of target gene fragments related to S100 A8 c DNA. The S100 A8 gene was successfully cloned and inserted into p LVX-IRES-puro. Conclusions： The recombinant lentiviral expression vector of human S100 A8 gene high expression is successfully constructed.