同济大学学报（医学版）2017，Vol.38Issue(6)：33-36,40,5.DOI:10.16118/j.1008-0392.2017.06.007
β-arrestin 2 shRNA慢病毒载体的构建及鉴定
Construction and verification of a lentivector for beta arrestin 2 short hairpin RNA
摘要
Abstract
Objective To construct and verify the β-arrestin 2 shRNA lentivector.Methods Four credible β-arrestin 2 knock-down sequences were searched.The linearized lentiviral vector pSicoR-GFP and shRNAs were constructed into aim plasmid pSicoR-shRNA under the effect of In-Fusion convertase.The aim plasmids were transfected into competent cells.The grown colonies were identified by colony PCR,and then the positive colonies were sequenced.Aligned recombinant lentivector plasmids and the other three helping plasmids were cotransfected into 293T cells by Lipofectamine 2000,and cell culture supernatant after 48 h of culture was collected to treat human nonsmall cell lung cancer cells A549,then the transfection efficiency was detected by qRT-PCR.Results β-arrestin shRNA lentivirus vector plasmid was constructed successfully,and the sequencing comparison results were consistent.qRT-PCR showed that one group of the β-arrestin 2 expression was only 56％ of the control group.Conclusion The β-arrestin 2 shRNA lentivector has been successfully constructed and its knock-down efficiency was verified in human non-small cell lung cancer cells A549.关键词
β-arrestin 2/shRNA/慢病毒/非小细胞肺癌Key words
β-arrestin 2/shRNA/lentivirus vector/non-small cell lung cancer分类
医药卫生引用本文复制引用
涂秋杰,任涛..β-arrestin 2 shRNA慢病毒载体的构建及鉴定[J].同济大学学报（医学版）,2017,38(6):33-36,40,5.基金项目
上海市浦东新区卫生系统重点学科建设项目(PWZx2014-10) （PWZx2014-10）
