医学信息2016，Vol.29Issue(29)：93-95,3.
人类microRNA-1246慢病毒抑制载体的构建以及鉴定
Construction and Identifacation of a Lentiviral Vector for MicroRNA-1246 of Human Inhibition
摘要
Abstract
Objective To construct and identify a lentiviral vector for microRNA-1246 inhibiton.Methods The gene sequences of microRNA-1246 Antisenseolignuclleotides were chosen, the antisense sequences of microRNA-1246 were designed and linked into linearized GV280 vector. The recombinants were screened and identified by colony PCR and DNA sequencing, The GV280-mir-246down,Helper1.0、Helper2.0 were mixed in suitable proportion and then transfected into the cell of 293T to get recombinant lentiviral vector.The concentrated and purified of virus bulk were detected the drops of it,indeed the virus was used to infect the carcinoma cell of Cervical squamous of siha .The GV280-mir-1246down,s expression in the siha cell tested by testing the GFP.Results ①The corrected of recombination lentiviral vector was screened from the bacterial which was anthenticated by PCR,DNA testing proved that the inserted gene sequence was correct . ②The cells of 293T which was transfected GV280-mir-1246down、Helper1、0 and Helper2.0 produced recombinant lentiviral vector. ③The target gene sequences were successfully inserted into siha cell by GV280 lentiviral vector ,the recombinant lentiviruses which carring mir-246down could infect and deliver mir-246down and GFP genes to siha cells. The infection efficiency was almost 100%. Conclusion The lentiviral microRNA inhibitor vector has been successfully constructed,which provides a stable tool for further study of therole of microRNA-1246 on cervical cancer.关键词
慢病毒载体/microRNA/抑制载体/宫颈癌/细胞增殖Key words
Lentiviral vector/MicroRNA/Inhibiton vector/Cervical cancer/Cell proliferation引用本文复制引用
赖月华,姚德生,杜萍,卢艳..人类microRNA-1246慢病毒抑制载体的构建以及鉴定[J].医学信息,2016,29(29):93-95,3.基金项目
国家自然科学基金项目资助(编号:8146039)广西自然科学基金 （编号:8146039）
