靶向XIAP的shRNA重组质粒的构建
X染色体连锁凋亡抑制蛋白;短发夹状RNA;肝癌;逆转录聚合酶链式反应;免疫蛋白印迹,吴小力,张琼,姚津剑,林菊生,张鹏,吴小力,通讯作者:,Constructionofrecombinantplasmidscontainingshorthairpi
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吴小力, 张琼, 姚津剑, 林菊生, 华中科技大学同济医学院附属同济医院消化内科 湖北省武汉市 430030
张鹏, 华中科技大学同济医学院附属同济医院普通外科 湖北省武汉市 430030
吴小力, 2000年华中科技大学同济医学院硕士, 主治医师, 主要从事消化系统疾病的研究.
通讯作者: 林菊生, 430030, 湖北省武汉市, 华中科技大学同济医学院附属同济医院消化内科. jslin@tjh.tjmu.edu.cn
收稿日期: 2007-04-26 修回日期: 2007-08-03
Construction of recombinant plasmids containing short hairpin RNA targeting the X-linked inhibitor of the apoptosis protein gene
Xiao-Li Wu, Peng Zhang, Qiong Zhang, Jin-Jian Yao, Ju-Sheng Lin
Xiao-Li Wu, Qiong Zhang, Jin-Jian Yao, Ju-Sheng Lin,Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Peng Zhang,Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Correspondence to:Ju-Sheng Lin, Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China. jslin@tjh.tjmu.edu.cn
Received:2007-04-26 Revised:2007-08-03
Abstract
AIM: To construct recombinant plasmids containing short hairpin RNA (shRNA) that targets the X-linked inhibitor of apoptosis protein (XIAP) gene, to assay the expression of XIAP in HepG2 cells after transfecting with recombinant plasmids, and to detect the RNAi effect of shRNA.
METHODS: Four pairs of shRNAs that target the XIAP gene were designed. The eukaryotic expression vector of psiRNA-Hhneo-XIAP was constructed and transfected via liposomes into HepG2 cells. XIAP mRNA expression was analyzed by reverse transcriptase-polymerase chain reaction. XIAP protein expression was examined by Western blotting. The difference in XIAP gene and protein expression levels between cells transfected or not transfected by psiRNA-Hhneo-XIAP was compared.
RESULTS: The successful construction of recombinant plasmids was confirmed by DNA sequencing of the inserted segments ......
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