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中国癌症防治杂志

中国癌症防治杂志 ›› 2013, Vol. 5 ›› Issue (2): 113-116.doi: 10.3969/j.issn.1674-5671.2013.02.07

• 基础研究 • 上一篇    下一篇

SATB1基因克隆及pEGFP-SATB1真核表达载体的构建和鉴定

  

  1. 广西医科大学附属肿瘤医院化疗三科；广西医科大学研究生学院
  • 出版日期:2013-06-25 发布日期:2013-06-27
  • 通讯作者: 岑 洪 E-mail:cen_hong@ 163.com
  • 基金资助:

    广西自然科学基金资助项目﹙桂科自0991230﹚

 Cloning of SATB1 cDNA and subcloning into a eukaryotic expression vector

  • Online:2013-06-25 Published:2013-06-27

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摘要： 目的 克隆特异AT序列结合蛋白1（special AT rich sequence binding protein，SATB1）基因全长编码序列，构建及鉴定其真核表达载体，为研究SATB1的功能及作用机制奠定实验基础。方法 以cDNA文库提供的pCMV6-XL6-SATB1为模板，通过PCR扩增SATB1基因全长编码序列，克隆入pGEM-T载体，再将SATB1基因插入真核表达载体pEGFP-N1，构建重组真核表达载体pEGFP-SATB1。结果 克隆的SATB1基因编码序列全长2 312 bp，经测序比对与Genbank登记的序列(BC001744.1)完全一致。经酶切及测序证实，SATB1基因正确插入pEGFP-N1载体中。结论 SATB1基因全长编码序列克隆正确，重组真核表达载体pEGFP-SATB1构建成功。

关键词:  , 特异AT序列结合蛋白1, 基因克隆, 真核表达载体

Abstract: Objective To clone the cDNA of the gene encoding special AT-rich sequence binding protein （SATB1） and to subclone it into eukaryotic expression vector pEGFP in order to investigate the function and mechanism of action of SATB1. Methods SATB1 cDNA was cloned from pCMV6-XL6-SATB1 by polymerase chain reaction （PCR） and then the target gene was subcloned into pGEM-T and subsequently into the eukaryotic expression vector pEGFP-N1 to give pEGFP-SATB1. Results The complete cDNA sequence of SATB1（2312 bp） was consistent with the reported sequence（Genbank BC001744.1）.Sequencing and restriction digest analysis re-vealed the target gene to be inserted in the expression vector in the correct orientation. Conclusion The SATB1 cDNA was success-fully cloned and used to construct recombinant eukaryotic expression vector pEGFP-SATB1.

Key words:  SATB1 gene, Gene cloning, Eukaryotic expression vector