Construction of the plasmid expressing gfp and MAD2 gene controlling separation of cell and its expression in HepG2 cell

SHI Qiong; et al

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Construction of the plasmid expressing gfp and MAD2 gene controlling separation of cell and its expression in HepG2 cell

更新时间：2024-01-17

- Construction of the plasmid expressing gfp and MAD2 gene controlling separation of cell and its expression in HepG2 cell
- Journal of Chongqing Medical University Issue 2, Pages: 180-182(2006)
- 作者机构：
  
  1. 重庆医科大学临床检验诊断学实验室
  2. 重庆医科大学临床检验诊断学实验室基础医学院核医学教研室
  3. ,重庆,400016
  4. 感染性疾病分子生物学教育部重点实验室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R346
- Published：2006，
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[1]施琼,翁亚光,徐远久,蒋洪彦,刘子杰,刘青松,蔡燕.细胞分裂调控基因MAD2与GFP融合的真核表达质粒的构建及表达[J].重庆医科大学学报,2006(02):180-182+239.

SHI Qiong, et al. Construction of the plasmid expressing gfp and MAD2 gene controlling separation of cell and its expression in HepG2 cell. [J]. Journal of Chongqing Medical University (2):180-182(2006)
[1]施琼,翁亚光,徐远久,蒋洪彦,刘子杰,刘青松,蔡燕.细胞分裂调控基因MAD2与GFP融合的真核表达质粒的构建及表达[J].重庆医科大学学报,2006(02):180-182+239. DOI：

SHI Qiong, et al. Construction of the plasmid expressing gfp and MAD2 gene controlling separation of cell and its expression in HepG2 cell. [J]. Journal of Chongqing Medical University (2):180-182(2006) DOI：

摘要

目的:构建MAD2基因与GFP融合的绿色荧光蛋白真核表达质粒

并在HepG2细胞中表达。方法:通过RT-PCR扩增

获得MAD2基因的完整序列。将此片断插入pEGFP-N1载体多克隆位点区

构建MAD2基因与GFP融合的绿色荧光蛋白真核表达质粒。应用脂质体转染技术转染HepG2细胞并观察表达的绿色荧光。结果:成功构建MAD2基因与GFP融合的绿色荧光蛋白真核表达质粒

并在HepG2细胞中观察到绿色荧光蛋白的表达。结论:该载体的成功构建

为研究MAD2基因在细胞分裂中的功能及定位建立了有效的观察体系。

Abstract

Objective: To establish an in vitro screening system for testing effects of MAD2 gene of embryonic cell genome.Methods: In order to construct the plasmid expressing MAD2-GFP fusion plasmid

the target gene fragment was obtained by RT-PCR amplification and inserted into pEGFP-N1 reporter vector.The structure of constructed plasmid was confirmed by electrophoresis analysis and DNA sequencing.The function of construct was confirmed by lipofectamine-mediated transient expression in HepG2 cells.Results: DNA sequencing showed that the inserted fragment of the constructed plasmid was the same as the template MAD2 genome.The HepG2 cells transfected with the constructed plasmid could express reporter gene of gfp.Conclusions: the plasmid expressing gfp gene controlled by MAD2 gene is successfully constructed and an in vitro testing system for evaluating founction of MAD2 gene in separation of cell is established.

关键词

MAD2基因绿色荧光蛋白HepG2细胞

Keywords

MAD2 genegreen fluorescence proteinHepG2 cell

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