Establishment and identification of siRNA eukaryotic expression vectors of recombinant targeting gene glucocorticoid receptor

LI Xiuying; et al

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Establishment and identification of siRNA eukaryotic expression vectors of recombinant targeting gene glucocorticoid receptor

更新时间：2024-01-17

- Establishment and identification of siRNA eukaryotic expression vectors of recombinant targeting gene glucocorticoid receptor
- Journal of Chongqing Medical University Issue 9, Pages: 943-945(2007)
- 作者机构：
  
  1. 重庆医科大学药学院药理教研室
  2. 云南克雷斯天然药物制药有限公司
  3. 第三军医大学大坪医院
- 作者简介：
- 基金信息：
- DOI：
  CLC： R346
- Published：2007，
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[1]李秀英,徐晓玉,孙锦帆,陈林,杨国汉.糖皮质激素受体GR真核表达载体的构建与鉴定[J].重庆医科大学学报,2007(09):943-945.

LI Xiuying, et al. Establishment and identification of siRNA eukaryotic expression vectors of recombinant targeting gene glucocorticoid receptor. [J]. Journal of Chongqing Medical University (9):943-945(2007)
[1]李秀英,徐晓玉,孙锦帆,陈林,杨国汉.糖皮质激素受体GR真核表达载体的构建与鉴定[J].重庆医科大学学报,2007(09):943-945. DOI：

LI Xiuying, et al. Establishment and identification of siRNA eukaryotic expression vectors of recombinant targeting gene glucocorticoid receptor. [J]. Journal of Chongqing Medical University (9):943-945(2007) DOI：

摘要

目的:构建糖皮质激素GR的真核表达载体

为探讨GR与中医肾阳虚的关系奠定基础。方法:以GR基因为靶基因

以pBS/U6-Neo质粒为载体

设计构建重组体

根据GR mRNA基因序列

设计有小发夹结构的2对寡核苷酸序列

克隆到空载体pBS/U6-Neo

转化DH5a菌株

提取质粒

进行PCR鉴定和测序分析。结果:经PCR鉴定筛出的重组体测序结果与目的序列相同

重组载体构建成功。结论:利用RNA干扰技术可成功构建小干扰RNA重组体

为后续稳定转染及培育转基因小鼠奠定基础。

Abstract

Objective:Establish eukaryotic expression vector of glucocorticoid receptor GR

to explore the relation of GR and tradition Chinese medicine.Methods:Recombinant were designed and established Supported by the National Natural Science Foundation of China(3047119)by targeting gene GR and plasmid pBS/U6-Neo based on GR mRNA sequence.Two pairs of oligonucleotides were synthesized and inserted into plasmid pBS/U6-Neo to generate siRNA eukaryotic expression vectors.DH5a strains were transformed.plasmids were extracted

and the recombinant sequences were identified by PCR and sequencing.Results:The result of recombinant sequence was the same as aim sequence.The recombinant vectors were established successfully.Conclusion:siRNA recombinant can be established successfully by RNAi technique

to use in further gene stability transfer and cultivate transgenic mouse.

关键词

糖皮质受体GRRNA干扰pBS/U6-Neo载体

Keywords

Glucocorticoid receptor(GR)RNA interferencepBS/U6-Neo vector

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Related Institution

Department of Radiology,The First Affiliated Hospital of Chongqing Medical University

Department of Emergency and Intensive Care Unit,The First Affiliated Hospital of Chongqing Medical University

Teaching and Research Section of Pathology,College of Basic Medicine,Molecular Medicine and Cancer Research Center,Chongqing Medical University

重庆医科大学公共卫生学院生殖医学教研室

Department of Biochemistry and Molecule Biology,College of Basic Medicine,Chongqing Medical University

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