IFI16基因反义干扰表达载体的构建与鉴定  

Construction and Identification of an Antisense-interference Expressing Vector for IFI16 gene

作　　者：张艳欧[1] 虞游[1] 赵文铖[1] 朱涵[1] 卢佳伟[1] 姚卉卉 陈建明[1] 

机构地区：[1]杭州师范大学生命与环境科学学院,浙江杭州310036

出　　处：《杭州师范大学学报（自然科学版）》2014年第1期42-45,共4页Journal of Hangzhou Normal University(Natural Science Edition)

基　　金：浙江省钱江人才计划项目(2010R10062)

摘　　要：为了构建可在人喉癌细胞中稳定表达IFI16基因短发夹RNA(shRNA)的表达载体,设计合成的IFI16基因shRNA片段,连接到经BamHI和EcoRI双酶切的pGreenPuroTMshRNA表达载体中,连接产物转化大肠杆菌后挑取几个抗性菌落,用PCR技术初步进行鉴定,经PCR初步鉴定为重组质粒的一个重组子DNA用测序进一步鉴定,测序结果显示成功构建了IFI16基因shRNA的表达载体pGreenPuro-IFI16shRNA.To construct the vector that can stably express IFI16 shRNA in human laryngocarcinoma cells ,the well designed and correctly synthesized IFI16 shRNA fragment was used to ligate into the pGreenPuro TM shRNA expressing vector that predigested with BamH I and EcoR I .Subsequently ,the recombinant vector was used to transform the E .coli and then the PCR was performed to identify the resistant colonies .Besides ,the identified recombinant DNA was subjected to sequence for the further identification .Finally ,the IFI16 shRNA expressing vector named pGreenPuro-IFI16 shRNA was correctly constructed as circumstantiated by the sequencing result .

关 键 词：IFI16基因 载体构建 

分 类 号：Q253[生物学—细胞生物学]