| 脯氨酰羟化酶RNA干扰慢病毒载体的构建与鉴定 |
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| 引用本文： | 马芳芳,王厚照,沈晓丽.脯氨酰羟化酶RNA干扰慢病毒载体的构建与鉴定[J].中国组织工程研究与临床康复,2011,15(41). |
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| 作者姓名： | 马芳芳 王厚照 沈晓丽 |
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| 作者单位： | 1. 解放军第174医院检验科,福建省厦门市,361003
2. 福建省立医院心血管病重点实验室,福建省福州市,350001 |
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| 摘 要： | 背景:慢病毒载体可稳定介导基因沉默且具有较高的转染效率.目的:构建并鉴定脯氨酰羟化酶 RNA 干扰慢病毒载体.方法:针对脯氨酰羟化酶2 基因序列设计RNA 干扰靶序列,合成靶序列的寡聚DNA,退火形成双链DNA,与经Age Ⅰ和EcoR Ⅰ酶切的pGCSIL-GFP 连接、转化大肠杆菌感受态细胞,产生重组RNA 干扰慢病毒表达载体,PCR 筛选阳性克隆,测序鉴定.结果与结论:PCR 和DNA 测序证实合成的含脯氨酰羟化酶短发卡RNA 慢病毒载体寡核苷酸链正确插入pGCSIL-GFP 载体.说明实验成功构建脯氨酰羟化酶基因RNA 干扰慢病毒载体.
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| 关 键 词： | 脯氨酰羟化酶 RNA 干扰 慢病毒 缺氧诱导因子1 克隆 |
Construction and identification of lentiviral vector of RNA interference of prolyl-4 hydroxylase-2 gene |
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| Ma Fang-fang,Wang Hou-zhao,Shen Xiao li.Construction and identification of lentiviral vector of RNA interference of prolyl-4 hydroxylase-2 gene[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2011,15(41). |
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| Authors: | Ma Fang-fang Wang Hou-zhao Shen Xiao li |
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| Institution: | Ma Fang-fang1,Wang Hou-zhao1,Shen Xiao li2 1Department of Laboratory Medicine,the 174 Hospital of Chinese PLA,Xiamen 361003,Fujian Province,China,2Key Laboratory of Cardiovascular Disease Fujian Provincial Hospital Fuzhou 350001 |
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| Abstract: | BACKGROUND:Lentiviral vector can stably mediate gene silencing with high transfection efficiency.OBJECTIVE:To construct and identify a lentiviral vector of RNA interference(RNAi) of prolyl-4 hydroxylase-2(P4HA2) gene.METHODS:The effective sequence of siRNA targeting P4HA2 gene was confirmed in our previous study.The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pGCSIL-GFP vector to construct a lentiviral vector which ex... |
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