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

XU Xue-mei; et al

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

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- Cloning of mouse signal transducers and activators of transcription-4/6 and construction of eukaryotic expression plasimid
- Journal of Chongqing Medical University Issue 8, Pages: 897-899(2008)
- 作者机构：
  
  重庆医科大学附属儿童医院呼吸科
- 作者简介：
- 基金信息：
- DOI：
  CLC： Q78
- Published：2008，
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[1]徐雪梅,符州,田代印,刘恩梅,王莉佳,黄英.小鼠STAT4、STAT6基因克隆及其真核表达质粒的构建[J].重庆医科大学学报,2008(08):897-899+910.

XU Xue-mei, et al. Cloning of mouse signal transducers and activators of transcription-4/6 and construction of eukaryotic expression plasimid. [J]. Journal of Chongqing Medical University (8):897-899(2008)
[1]徐雪梅,符州,田代印,刘恩梅,王莉佳,黄英.小鼠STAT4、STAT6基因克隆及其真核表达质粒的构建[J].重庆医科大学学报,2008(08):897-899+910. DOI：

XU Xue-mei, et al. Cloning of mouse signal transducers and activators of transcription-4/6 and construction of eukaryotic expression plasimid. [J]. Journal of Chongqing Medical University (8):897-899(2008) DOI：

摘要

目的:分别克隆小鼠信号传导及转录活化因子-4（Signal transducers and activators of transcription-4

STAT4）、信号传导及转录活化因子-6（Signal transducers and activators of transcription-6

STAT6）基因全长cDNA序列

构建并鉴定其真核表达质粒pIRES2-EGFP-STAT4/STAT6。方法:采用RT-PCR方法从BALB/c小鼠脾脏组织中扩增STAT4/STAT6基因全长cDNA

T/A克隆到pMD19-T载体中

双酶切后将cDNA亚克隆入pIRES2-EGFP载体

构建pIRES2-EGFP-STAT4/STAT6质粒

经限制性内切酶鉴定并测序。结果:小鼠STAT4/STAT6基因cDNA正确克隆入pIRES2-EGFP表达载体

与Genebank中STAT4/STAT6基因序列一致。结论:成功构建pIRES2-EGFP-STAT4/STAT6表达质粒

为进一步研究STAT4/STAT6蛋白表达和相互作用奠定了基础。

Abstract

Objective:To clone mouse signal transducers and activators of transcription(STAT) 4 and STAT6 gene and construct a eukaryotic expression plasimid.Methods:The cDNA of mouse STAT4 and STAT6 gene amplified with RT-PCR from normal mouse spleen tissue were cloned into pMD19-T vector by T/A ligation.After double digested

STAT4/STAT6 cDNA fragment was subcloned into pIRES2-EGFP vector to construct a eukaryotic expression plasimid.Restriction endonucleases analysis and sequencing were used to comform the recombinant plasimid.Results:The full length STAT4/STAT6 cDNA was correctly inserted into eukaryotic expression plasimid

and its sequencing was consistent with reported sequence derived from Genebank.Conclusion:The successful construction of eukaryotic expression plasimid provides a basis for futher studies on the infection of STAT4/STAT6 interaction to the downstream cytokines.

关键词

哮喘信号传导及转录活化因子克隆真核表达质粒

Keywords

asthmaSignal transducers and activators of transcriptioncloneEukaryotic expression plasimid

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Related Institution

Department of Respiratory Medicine，Children’s Hospital，Zhejiang University School of Medicine/ National Clinical Research Center for Child Health

Department of Pediatrics，The Affiliated Hospital of Nanjing University of Chinese Medicine，The First Clinical Medical College of Nanjing University of Chinese Medicine

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Department of Emergency and Critical Care Medicine,The First Affiliated Hospital of Chongqing Medical University

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