Cloning of CGI-100 gene and reconstruction of its eukaryotic expressive vector

HUANG Fengxia; et al

您当前的位置：

首页 >

文章列表页 >

Cloning of CGI-100 gene and reconstruction of its eukaryotic expressive vector

更新时间：2024-01-17

- Cloning of CGI-100 gene and reconstruction of its eukaryotic expressive vector
- Journal of Chongqing Medical University Issue 8, Pages: 785-788(2007)
- 作者机构：
  
  1. 重庆医科大学临床生化教研室临床检验诊断学重庆市重点实验室
  2. 重庆医科大学临床生化教研室临床检验诊断学重庆市重点实验室,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC： R73-3
- Published：2007，
扫描看全文
[1]黄凤霞,张彦,王伟佳.未知基因CGI-100的克隆及其真核表达载体的构建[J].重庆医科大学学报,2007(08):785-788.

HUANG Fengxia, et al. Cloning of CGI-100 gene and reconstruction of its eukaryotic expressive vector. [J]. Journal of Chongqing Medical University (8):785-788(2007)
[1]黄凤霞,张彦,王伟佳.未知基因CGI-100的克隆及其真核表达载体的构建[J].重庆医科大学学报,2007(08):785-788. DOI：

HUANG Fengxia, et al. Cloning of CGI-100 gene and reconstruction of its eukaryotic expressive vector. [J]. Journal of Chongqing Medical University (8):785-788(2007) DOI：

摘要

目的:为探讨研究未知基因CGI-100的功能

克隆人CGI-100基因并构建其真核表达载体。方法:通过RT-PCR从人白血病K562细胞中克隆CGI-100基因全长编码区

将该cDNA片段亚克隆至载体pMD18-T SimpleT中

经测序鉴定后

用BamHⅠ和PstⅠ双酶切将目的cDNA片段定向克隆至相同处理的pIRES2-EGFP真核表达质粒中

然后经酶切、PCR鉴定和DNA序列测定三重鉴定方法对重组质粒进行鉴定。结果:经酶切、PCR及测序鉴定证实

CGI-100基因全长编码区cDNA被准确正向插入至pIRES2-EGFP质粒的酶切位点BamHⅠ和PstⅠ之间

未发现碱基突变或移位。结论:成功地构建并鉴定了含CGI-100基因全长编码区的真核表达质粒

为以后研究该基因的功能奠定了基础。

Abstract

Objective:To clone human CGI-100 gene and reconstruct its eukaryotic expressive vector for further investigation.Methods:The full coding domain sequence of human CGI-100 gene was cloned from human leukemic K562 cells with RT-PCR and was sub-cloned into pMD18-T Simple vector.After confirmed by DNA sequencing

the targeted DNA fragment

digested with BamHⅠand PstⅠ

was directionally cloned into eukaryotic expressive plasmid pIRES2-EGFP

then

the reconstructed plasmid was identified with enzyme digestion

PCR and sequencing.Results:It was demonstrated that the full coding domain sequence of human CGI-100 gene was accurately cloned into digestion sites between BamHⅠand PstⅠin pIRES2-EGFP without mutation and transposition.Conclusion:The reconstruction and verifying of eukaryotic expressive plasmid containing CGI-100 gene are successful

which establishes the foundation of further investigation.

关键词

CGI-100基因真核表达载体克隆

Keywords

CGI-100 geneEukaryotic expressive vectorclone

references

Views

101

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

A method for constructing extensive mutagenesis of HBV polymerase

Cloning,expression and purification of human canstatin and assay of its bioactivity

Expression,identification and purification of the resuscitation-promoting factor B of Mycobacterium tuberculosis

Expression,identification and purification of the resuscitation-promoting factor A of mycobacterium tuberculosis

Cloning of mouse signal transducers and activators of transcription-4/6 and construction of eukaryotic expression plasimid

Related Author

Hu Jieli

Huang Ailong

Zhang Wenlu

Yang Liuqing

Liu Ya

曾昭淳

et al

ZHANG Xiangrong

Related Institution

Key Laboratory of Molecular Biology on Infectious Diseases Founded by Ministry of Education,Chongqing Medical University

The Department of Biochemistry,Academy of Basic Medicine,Chongqing Medical University

重庆医科大学基础医学院生物化学教研室

Department of Immunology,Chongqing Medical University

重庆医科大学附属第一医院眼科

⁰