重庆医学Issue(1)：73-75,83,4.DOI:10.3969/j.issn.1671-8348.2015.01.026
慢病毒载体靶向介导p66shc基因沉默
Study on lentiviral vector target inducing p66 shc gene silencing
摘要
Abstract
Objective To construct p66shc gene interfering lentivirus vectors recombination and transfect it to 293T cells ,RNA interfering was carried out to induce p66shc gene silence ,so as to provide basis for further study of the p66shc function .Methods Screening of three RNA targets which were named after p66shc‐shc1 ,p66shc‐shc2 ,p66shc‐shc3 ,cloned into the pLenR‐GPH vec‐tor ,which contained green fluorescent protein(GFP) and transformed into DH5αcells .The positive clone were picked out for right sequencing and transfected to 293T cells with pRsv‐REV ,pMDlg‐pRRE ,pMD2G .The expression of GFP in inverted fluorescence microscope confirmed the virus packaging success .Fluorescence quantitative PCR and Western blot technology were used to investi‐gate the expression of p66shc at the molecular and protein levels ,p66shc‐shc1 target of effective silencing p66shc gene was selected to prepare for subsequent tests .Results The shRNA lentivirus vector was constructed which could express p66shc and was trans‐fected into 293T cells successfully .Fluorescence quantitative PCR and Western blot technology were used to investigate p66shc gene silence by RNA interference .Conclusion The lentivirus RNAi vector of targeted expression p66shc could induce p66shc gene si‐lence at the molecular and protein levels after transfected into 293T cells by RNA interference .关键词
慢病毒属/RNA干扰/p66shcKey words
lentiviral vector/RNA interference/p66shc分类
社会科学引用本文复制引用
张婵,董文斌,赵帅,李清平,康兰,雷小平,郭琳,翟雪松..慢病毒载体靶向介导p66shc基因沉默[J].重庆医学,2015,(1):73-75,83,4.基金项目
中华儿科杂志第二届双鹤珂立苏科研基金；四川省教育厅科研基金（08ZA150）；四川省卫生厅科研基金（90191）。 （）
