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| Smad3基因小干扰RNA的序列设计及其表达载体的构建 | |
| 引用本文： | 王泽荣,王建浩,胡从莉,沈秀娟,吴妹英,吴士良. Smad3基因小干扰RNA的序列设计及其表达载体的构建[J]. 实用肝脏病杂志, 2010, 13(5): 328-330,368. DOI: 10.3969/j.issn.1672-5069.2010.05.004 |
| 作者姓名： | 王泽荣 王建浩 胡从莉 沈秀娟 吴妹英 吴士良 |
| 作者单位： | 1. 215007,江苏省苏州市第五人民医院;苏州大学基础医学与生物科学学院 2. 苏州大学基础医学与生物科学学院 3. 江苏省苏州市第五人民医院,215007 |
| 摘 要： | 目的构建大鼠Smad3基因小干扰RNA的表达质粒,为肝纤维化RNAi介导的基因治疗打下基础。方法利用互联网资源针对Smad3基因设计并合成三条可能的小干扰RNA,脂质体瞬时转染大鼠肝星状细胞,RT-PCR检测Smad3基因的表达情况。然后将这三条小干扰RNA插入到RNA干扰载体pRNAT-U6.1/Neo中,构建成三条干扰载体并测序。结果 RT-PCR和测序结果表明,成功构建了Smad3基因的RNA干扰真核表达载体。结论重组质粒pRNAT-SiSmad3的成功构建,为后续的Smad3基因下调实验提供了可能。 |
| 关 键 词： | 肝纤维化 Smad3 干扰载体,siRNA 肝星状细胞 |
Designment and vector construction of siRNA sequences to Smad3 gene in vitro |
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| WANG Zerong,WANG Jianhao,HU Congli,et al.. Designment and vector construction of siRNA sequences to Smad3 gene in vitro[J]. Journal of Clinical Hepatology, 2010, 13(5): 328-330,368. DOI: 10.3969/j.issn.1672-5069.2010.05.004 | |
| Authors: | WANG Zerong WANG Jianhao HU Congli et al. |
| Affiliation: | WANG Zerong,WANG Jianhao,HU Congli,et al.Department of Biochemistry,Medical School,Suzhou University,Suzhou 215123,Jiangsu Province,China |
| Abstract: | Objective To construct an expression vector with small interfering RNAs of Smad3 in vitro.Methods Three possible small interfering RNAs to Smad3 gene were designed and synthesized,subsequently transfected into rat hepatic stellate cells using Liposome 2000.RT-PCR was used to detect the expression of the Smad3 gene.Then the strands were cloned into pRNAT-U6.1/Neo vector to construct the recombinant plasmids and confirmed by sequencing.Results Both RT-PCR and sequencing assays showed that the recombinant vect... |
| Keywords: | Smad3 |
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