靶向遗传印记基因PEG10 的siRNA 真核表达载体的构建及鉴定

The Construction and Identification of siRNA Eukaryotic Expression Vector Targeting Genetic Imprinted Gene PEG10

摘要: 目的构建并鉴定针对遗传印记基因PEG10的siRNA真核表达载体。方法根据PEG10基因cDNA序列，设计针对目的基因的4个siRNA靶序列，将其插入H1启动子下游，克隆到真核表达载体psiRNA-hHlneo中，通过酶切鉴定和DNA测序鉴定。结果酶切鉴定及DNA测序结果显示插入片断正确。结论本研究成功构建针对PEG10的真核表达载体，可能成为肝癌基因治疗中的一种新型、高效的治疗载体。

Abstract: Objective 　To construct and identify the siRNA eukaryotic expression vector targeting PEG10. Methods 　Four siRNAs were designed according to the coding sequence of PEG10 gene, and cloned into the downst ream of H1 promoter of psiRNA-hH1neo. The const ructed recombinant was analyzed and identified by Ase Ⅰendonuclease digestion and DNA sequencing. Results 　The constructed psiRNA plasmid digested with Ase Ⅰwas linearized. The sequencing result confirmed that the sequence of inserted fragment was correct . Conclusion 　Eukaryotic expression vector of siRNA targeting PEG10 gene was successfully constructed, and should be a novel effective expression vector for HCC gene therapy.