基因表达__(期刊论文)

反义cortactin基因真核重组质粒的构建及鉴定

http://www.100md.com 《第四军医大学学报》 2006年第8期

基因表达,,基因表达；高转移肝癌细胞MHCC97；逆转录聚合酶链反应；人皮层肌动蛋白cortactin,0引言,1材料和方法,2结果,3讨论

     Construction and identification of recombinant eukaryotic expression vector of human antisense cortical actinassociated protein gene

    XU ZhuDing, SONG ZhenShun, AN JiaZe, LI Ren, ZHANG FuQin, DOU KeFeng

    Department of Hepatobiliary Surgery, Xijing Hospital, Fourth Military Medical University, Xian 710033, China

    【Abstract】 AIM: To construct a recombinant eukaryotic expression vectorpcDNA3.0/cortactin containing functional region of human antisense cortical actinassociated protein gene (cortactin), so as to provide a tool for the further study on the impact of cortactin in hepatocellular carcinoma (HCC) metastasis. METHODS: Total RNA was extracted by using Trizol onestep method from cell line MHCC97. CDNA was amplified using reverse transcriptasepolymerase chain reaction (RTPCR). Product of PCR, amplified by being transformed into E.coli Dh5α, was inserted reversely into the eukaryotic expression vector after digestion and ligation using restriction endonucleases and ligase. RESULTS: The correct cortactin cDNA clone was obtained and it was confirmed that cortactin cDNA was inserted reversely into the eukaryotic expression vector correctly using PCR, digestion identification and sequencing. CONCLUSION: The recombinant eukaryotic expression vector pcDNA3.0/cortactin is successfully constructed. ......

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