靶向人SSH1L的siRNA真核表达载体的构建及鉴定  

Construction and identification of eukaryotic expression vector of SSH1L-targeted siRNA

作　　者：邢志军[1] 赵建武[1] 王岩[2] 高忠礼[1] 

机构地区：[1]吉林大学中日联谊医院,吉林长春130033 [2]吉林大学病理生物学教育部重点实验室,吉林长春130021

出　　处：《中国实验诊断学》2009年第1期12-14,共3页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科技厅白求恩专项基金(No200705295);吉林省省科技厅国际合作项目(No20080723);国家自然科学基金资助项目(No30772488)

摘　　要：目的构建靶向人SSH1L的siRNA真核表达载体(pSUPER-SSH1L)。方法将设计好的SSH1L的siRNA的寡聚脱氧核苷酸链与真核表达载体pSUPER连接,构建重组pSUPER-SSH1L真核表达载体。对该重组表达载体进行酶切鉴定及DNA测序;通过Western blot检测SSH1L蛋白的表达。结果酶切鉴定、DNA测序以及Western blot检测结果证实表达质粒构建成功,无碱基突变。结论成功构建了靶向人SSH1L基因表达的siRNA干扰质粒载体pSUPER-SSH1L,为进一步运用RNA干扰技术进行SSH1L基因功能研究奠定了基础。Objective To construct a recombinant eukaryotic expression piasmid carrying human SSH1L-siRNA gene.Methons To connect the designed siRNA ＇ oligodeoxynucleofide chain of SSH1L with eukaryofic expression vector pSUPER, construct a recombinant eukaryotic expression plasmid pSUPER- SSH1L. Then to identify the recombinant expression plasmid by enzyme analysis and carry out DNA sequencing;and to check the expression of SSH1L protein through Western blot.Results The result of enzyme analysis,DNA sequencing and Western blot confirmed that the eukaryotic expression vector pSUPER-SSH1L was successfully con- structed,no basic radical mutation. Conclusion Eukaryotie expression vector pSUPER-SSH1L was successfully constructed, which was the basis for further studying SSH1L gene＇funcfion operating RNAi technique.

关 键 词：SSH1L SIRNA 真核表达载体 

分 类 号：R346[医药卫生—基础医学]