Construction and identification of VEGF shRNA eukaryotic expression plasmid in rat

FEI Chun-mei; et al

doi:10.13406/j.cnki.cyxb.2010.02.039

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Construction and identification of VEGF shRNA eukaryotic expression plasmid in rat

更新时间：2024-01-17

- Construction and identification of VEGF shRNA eukaryotic expression plasmid in rat
- Journal of Chongqing Medical University Vol. 35, Issue 2, Pages: 206-209(2010)
- 作者机构：
  
  重庆医科大学附属第一医院眼科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.02.039
  CLC： R346;R772
- Published：2010，
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[1]费春梅,周善璧.大鼠血管内皮生长因子shRNA表达质粒的构建及鉴定[J].重庆医科大学学报,2010,35(02):206-209.

FEI Chun-mei, et al. Construction and identification of VEGF shRNA eukaryotic expression plasmid in rat. [J]. Journal of Chongqing Medical University 35(2):206-209(2010)
[1]费春梅,周善璧.大鼠血管内皮生长因子shRNA表达质粒的构建及鉴定[J].重庆医科大学学报,2010,35(02):206-209. DOI： 10.13406/j.cnki.cyxb.2010.02.039.

FEI Chun-mei, et al. Construction and identification of VEGF shRNA eukaryotic expression plasmid in rat. [J]. Journal of Chongqing Medical University 35(2):206-209(2010) DOI： 10.13406/j.cnki.cyxb.2010.02.039.

摘要

目的:构建大鼠血管内皮生长因子（Vascular endothelial growth factor

VEGF）短发夹RNA（Short hairpin RNA

shRNA）真核表达质粒

为利用RNA干扰技术（RNA interference

RNAi）从转录后水平抑制角膜新生血管的研究做准备。方法:用DNA重组技术根据大鼠VEGF mRNA序列设计并合成3条shRNA寡核苷酸片段

退火形成双链并克隆进入载体pSilencer2.0 U6中

进行酶切鉴定和测序。结果:酶切证明构建的shRNA序列已插入载体

经测序证明与设计的相同

获得大鼠VEGF shRNA表达质粒。结论:成功构建了3条大鼠VEGF shRNA真核表达质粒

为靶向VEGF的RNAi抑制角膜新生血管奠定了基础。

Abstract

Objective:To construct rat vascular endothelial growth factor short hairpin RNA eukaryotic plasmid expression vector

in order to provide a basis for inhibition of corneal neovascularization by RNA interference.Methods:Three different shRNAs targeting the coding sequence of the rat VEGF mRNA were designed.After annealing the formed double stranded DNAs were inserted into the plasmid pSilencer2.0 U6.Then the recombinant plasmids were identified by enzyme digestion and DNA sequencing.Results:The three VEGF shRNAs were successfully inserted into the plasmid vector

and the coding sequences of the obtained shRNAs were consistent with the designed fragments.Conclusion:The recombinant plasmids of VEGF shRNAs were successfully constructed

which will be useful to inhibit corneal neovascularization.

关键词

血管内皮生长因子RNA干扰质粒shRNA

Keywords

VEGFRNA interferenceplasmidShort hairpin RNA

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