湖北医药学院学报2011，Vol.30Issue(2)：109-111,115,4.
Skp2-siRNA重组腺病毒载体构建及鉴定
Construction and Identification of Recombinant Adenovirus Vector of siRNA Targeting Skp2
摘要
Abstract
Objective To construct an edenovirus vector expressing small interfering RNA(siRNA) targeting to Skp2. Methods The siRNA containing DNA sequence targeting to Skp2 and its negative control sequence were designed,synthesized,annealed and subcloned into pShuttle-H1 containing the green fluorescent protein. The linearized shuttle plasmid by Pme I was recombined with back-bone pAdEasy-1 in BJ5183 bacteria. The recombinant pAd-Skp2-siRNA adenovirus particles were produced by transfection of AD293 cells and subsequently infected colonic cancer SW480 cells. The Skp2 protein levels were detected by Western blot. Results The adenovirus vectors were verified by enzyme digestion and DNA sequencing,the infection efficiency of constructed adenovirus was high,and could obviously inhibit the expression of Skp2. Conclusion The recombinant pAd-Skp2-siRNA and negative control bacterial were successfully constructed by bacterial homologous recombination.关键词
Skp2/siRNA/腺病毒载体分类
生物科学引用本文复制引用
魏刚,孙泽群,孔霞,黄永章,王斌,唐俊明,徐少勇..Skp2-siRNA重组腺病毒载体构建及鉴定[J].湖北医药学院学报,2011,30(2):109-111,115,4.基金项目
湖北省教育厅科研基金(200524001). （200524001）
