| VEGF靶向性siRNA表达载体的构建及其沉寂作用 |
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| 引用本文： | 李玲,王风华,连鸿凯,白玉,蔡朝阳,赵国强,马俊芬. VEGF靶向性siRNA表达载体的构建及其沉寂作用[J]. 医学争鸣, 2009, 30(21): 2332-2334 |
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| 作者姓名： | 李玲 王风华 连鸿凯 白玉 蔡朝阳 赵国强 马俊芬 |
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| 作者单位： | 郑州市骨科医院,郑州大学,河南,郑州,450052;郑州大学基础医学院,河南,郑州,450052;郑州大学第五附属医院,河南,郑州,450052;郑州市骨科医院,郑州大学,河南,郑州,450052;郑州大学基础医学院,河南,郑州,450052 |
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| 基金项目： | 郑州市科技攻关项目(20061952) |
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| 摘 要： | 目的：构建针对VEGF的siRNA表达载体,转染入人骨肉瘤细胞系MG-63观察其对细胞VEGF表达的沉默效应.方法：设计VEGF发卡样siRNA,依据设计合成两条互补的寡核苷酸链,退火后连接入T载体,转化扩增后进行序列测定.用脂质体包裹转染骨肉瘤细胞MG-63,用RT-PCR方法检测细胞VEGF mRNA表达水平.结果：把针对VEGF基因的siRNA的双链寡合苷酸片断克隆到表达载体pSuperneo,经过酶切鉴定与测序,结果正确;稳定转染人骨肉瘤细胞系MG-63后,用RT-PCR方法检测细胞VEGF mRNA表达水平明显降低.结论：成功构建出针对VEGF的siRNA表达载体pSuper-neo-VEGF,转染骨肉瘤细胞后能显著降低细胞VEGF mRNA表达.
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| 关 键 词： | RAN干扰 血管内皮生长因子 骨肉瘤 |
Vector construction and silencing effect of VEGF gene targeted small interfering RNA |
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| LI Ling,WANG Feng-Hua,LIAN Hong-Kai,BAI Yu,CAI Chao-Yang,ZHAO Guo-Qiang,MA Jun-FenOrthopedic Hospital of Zhengzhou,School of Basic Medical Science,Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou ,China. Vector construction and silencing effect of VEGF gene targeted small interfering RNA[J]. Negative, 2009, 30(21): 2332-2334 |
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| Authors: | LI Ling WANG Feng-Hua LIAN Hong-Kai BAI Yu CAI Chao-Yang ZHAO Guo-Qiang MA Jun-FenOrthopedic Hospital of Zhengzhou School of Basic Medical Science Fifth Affiliated Hospital Zhengzhou University Zhengzhou China |
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| Affiliation: | LI Ling1,2,WANG Feng-Hua3,LIAN Hong-Kai1,BAI Yu1,CAI Chao-Yang1,ZHAO Guo-Qiang2,MA Jun-Fen21Orthopedic Hospital of Zhengzhou,2School of Basic Medical Science,3Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052,China |
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| Abstract: | AIM:To construct VEGF gene-targeted small interfering RNA(siRNA) and its expression vector; and to observe the expression level of its expression vector VEGF gene in human osteosarcoma MG-63 cell transfected by the vector. METHODS:First to design VEGF gene-targeted hairpin siRNA,then to synthesize two complementary oligo nucleotide strand,after annealing,to insert the two-strand oligo nucleotide into pSuper neo vetor,which was then restrictive enzyme digested and sequenced. After that we transfected human o... |
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| Keywords: | siRNA VEGF osteosarma |
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