| 人血管内皮生长因子mRNA靶向siRNA表达载体的构建和表达 |
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| 引用本文： | 丁培杰,杨书钦,曹明瑞,臧文巧,陈宗德,赵国强. 人血管内皮生长因子mRNA靶向siRNA表达载体的构建和表达[J]. 郑州大学学报(医学版), 2007, 42(2): 231-233 |
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| 作者姓名： | 丁培杰 杨书钦 曹明瑞 臧文巧 陈宗德 赵国强 |
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| 作者单位： | 漯河医学高等专科学校微生物学与免疫学教研室,漯河,446200;河南省煤田地质局-队医院,新郑,451150;郑州大学第一附属医院妇产科,郑州,450052;郑州大学基础医学院微生物学与免疫学教研室,郑州,450001 |
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| 基金项目： | 教育部211工程重点学科建设项目 |
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| 摘 要： | 目的:构建人血管内皮生长因子(VEGF)mRNA靶向siRNA表达载体,观察其表达情况.方法:设计人VEGF靶向的发夹样siRNA,依据设计,合成2条互补的寡核苷酸链,退火后连接入pSUPERneo-GFP载体,转化扩增后得到重组载体pSUPERneo-GFP-siVEGF,进行序列测定.用脂质体转染法将重组载体转染食管癌细胞株Eca109,采用RT-PCR检测转染细胞VEGF mRNA的表达水平. 结果:经过酶切鉴定与测序,pSUPERneo-GFP-siVEGF构建成功.稳定转染该重组体的Eca109细胞VEGF mRNA表达水平明显降低.结论:成功构建了针对VEGF mRNA的siRNA载体, 转染细胞后可显著抑制VEGF mRNA的表达.
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| 关 键 词： | RNA干扰 VEGF 表达 |
| 收稿时间： | 2006-05-19 |
| 修稿时间： | 2006-05-19 |
Vector construction and silencing effect of VEGF gene targeted small interfering RNA |
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| DING Peijie,YANG Shuqin,CAO Mingrui,ZANG Wenqiao,CHEN Zongde,ZHAO Guoqiang. Vector construction and silencing effect of VEGF gene targeted small interfering RNA[J]. Journal of Zhengzhou University: Med Sci, 2007, 42(2): 231-233 |
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| Authors: | DING Peijie YANG Shuqin CAO Mingrui ZANG Wenqiao CHEN Zongde ZHAO Guoqiang |
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| Affiliation: | 1. Department of Microbiology and Immunology, Luohe Medical College, Luohe 4462000; 2 . Hospital of Henan Province Coalfield Geological Bureau Team 1, Xinzheng 451150; 3. Department of Gynecology and Obstetrics, the First Affiliated Hospital, Zhengzhou Unwersity, Zhengzhou 450052 ;4.College of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450052 |
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| Abstract: | Aim: To construct VEGF gene-targeted small interfering RNA(siRNA) vector. Methods: VEGF gene-targeted hairpin siRNA was designed, then two complementary oligo nucleotide strand were synthesized and inserted into pSUPERneo-GFP vector after annealing to get the new vector pSUPERneo-GFP-siVEGF, which was digested by restrictive enzyme and sequenced. After that, the new vector was transfected into human esophageal carcinoma Eca109 cell using lipofectamine method, and the mRNA exprssion level of VEGF gene in cells was detected by RT-PCR.Results and Conclusion: pSUPERneo-GFP-siVEGF is built successfully,and the expression level of VEGF mRNA in transfected Eca109 was decreased. |
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| Keywords: | RNA interference VEGF expression |
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