Construction of pEGFP-VEGF recombinant plasmid and its expression in osteoblast

FAN Wei; et al

doi:10.13406/j.cnki.cyxb.2005.01.008

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Construction of pEGFP-VEGF recombinant plasmid and its expression in osteoblast

更新时间：2024-01-17

- Construction of pEGFP-VEGF recombinant plasmid and its expression in osteoblast
- Journal of Chongqing Medical University Issue 1, Pages: 27-29(2005)
- 作者机构：
  
  1. 重庆医科大学临床学院骨科
  2. 重庆医科大学临床学院骨科,重庆,400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2005.01.008
  CLC： R68
- Published：2005，
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[1]范伟,安洪,蒋电明.pEGFP-VEGF重组质粒的构建及其在成骨细胞中的表达[J].重庆医科大学学报,2005(01):27-29+38.

FAN Wei, et al. Construction of pEGFP-VEGF recombinant plasmid and its expression in osteoblast. [J]. Journal of Chongqing Medical University (1):27-29(2005)
[1]范伟,安洪,蒋电明.pEGFP-VEGF重组质粒的构建及其在成骨细胞中的表达[J].重庆医科大学学报,2005(01):27-29+38. DOI： 10.13406/j.cnki.cyxb.2005.01.008.

FAN Wei, et al. Construction of pEGFP-VEGF recombinant plasmid and its expression in osteoblast. [J]. Journal of Chongqing Medical University (1):27-29(2005) DOI： 10.13406/j.cnki.cyxb.2005.01.008.

摘要

目的构建一种含人血管内皮细胞生长因子121（VEGF121）的质粒

以绿色荧光蛋白作为报告基因

观察其在成骨细胞中的表达。方法从人胎脑文库中

以PCR方法扩增出人VEGF121基因

构建入增强绿色荧光蛋白表达质粒pEGFP-C1

脂质体法转染成骨细胞

荧光显微镜下观测绿色荧光蛋白的表达;免疫细胞化学及ELISA检测VEGF蛋白的表达。结果PCR扩增出人VEGF121基因;构建入pEGFP-C1

体外成功转染入成骨细胞

在荧光显微镜下可见强绿色荧光蛋白的表达

免疫细胞化学及ELISA检测到VEGF蛋白的表达。结论重组质粒pEGFP-VEGF体外转染入成骨细胞后

目的基因能够在细胞有效表达

检测方法简便可靠

为利用转染该种目的基因的成骨细胞进行下一步实验提供了理论支持。

Abstract

Objective:To construct a vector containing human vascular endothelial growth factor

VEGF 121 (VEGF 121 ) with green fluorescence protein as the reporter gene

and then to observe its expression in osteoblast.Methods:VEGF 121 gene was amplified from foetus brain library by PCR

then ligated with pEGFP-C1 to form recombinant plasmid pEGFP-VEGF and transfected into osteoblast by liposome.The green fluorescence protein expression was observed by fluorescence microscopy and the VEGF expression was last detected by immunocytochemistry and ELISA.Results:VEGF 121 gene fragment was amplified by PCR and ligated to pEGFP-C1

then transfected into osteoblast successfully.The strong expression of green fluorecensce protein was observed by fluorecensce microscopy

and the expression of VEGF protein was detected by immunocytochemistry and ELISA.Conclusion:Recombinant plasmid pEGFP-VEGF is effectivly expressed after being transfected into osteoblast in vitro

which provides theoretical support for further study with its simple and reliable detecting methods.

关键词

血管内皮细胞生长因子质粒构建基因转染成骨细胞

Keywords

VEGFPlasmid constructiongene transfectionosteoblast

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