昆明医科大学学报Issue(10)：58-61,72,5.
pSIREN-HIF-1α/shRNA表达载体的构建与鉴定
Construction and Identification of pSIREN-HIF-1α/shRNA Expression Vector
摘要
Abstract
Objective To construct and identity pSIREN-HIF-1α/shRNA expression vector in order to make foundation of gene therapy for further exploration of RNA interference to nasopharyngeal darcinoma. Methods According to HIF-1αcDNA gene sequence in the gene bank (NM_001530/NM_181054), a pair of 60 nt oligonucleotides each containing the sites of restriction endonuclease at both ends,were designed and synthesized by Reynolds design principles. Oligonucleotides were annealed and ligated with linedrized RNAi-Ready pSIREN-RetroQ-ZsGreen.Transfected into JM109, the recombinants were finally sequenced and identified by 1%agarose gel electrophoresis. Results The size of the target gene fragment amplified by PCR was 470 bp and in accordance with the expected result.pSIREN-HIF-1α was successfully constructed and identitfied by 1%agarose gel electrophoresis.Sequence analysis of inserted fragment revealed the same sequence as synthesized shRNA Oligonucleotides. Conclusion pSIREN-HIF-1α /shRNA expression vector has been successfully constructed, and can make the foundation of research using liposome packaging transfectiing nasopharyngeal darcinoma cell for the next step .关键词
RNA干扰/鼻咽癌/质粒/HIF-1α/短发卡RNAKey words
RNA interference/Nasopharyngeal darcinoma/Plasmids/HIF-1α/shRNA分类
医药卫生引用本文复制引用
赵留芳,张书萍,李晓江,任艳鑫,张康,喻博,马静,隋军..pSIREN-HIF-1α/shRNA表达载体的构建与鉴定[J].昆明医科大学学报,2013,(10):58-61,72,5.基金项目
云南省卫生科技计划项目 （）
