The expression of open reading frame of unknown KH gene from K562 cells in E.coli

HE Yujuan; et al

doi:10.13406/j.cnki.cyxb.2004.02.002

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The expression of open reading frame of unknown KH gene from K562 cells in E.coli

更新时间：2024-01-17

- The expression of open reading frame of unknown KH gene from K562 cells in E.coli
- Journal of Chongqing Medical University Issue 2, Pages: 124-126(2004)
- 作者机构：
  
  1. 重庆医科大学基础医学院临床生化教研室
  2. 重庆市永川市人民医院
  3. 重庆医科大学临床学院血液科
  4. 重庆医科大学基础医学院临床生化教研室,重庆,400016
  5. ,重庆,402160
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2004.02.002
  CLC： Q781
- Published：2004，
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[1]何於娟,欧一衡,张彦,刘小珊,马凌娣,涂植光,蒋纪恺.K562细胞未知基因KH的开放阅读框架在大肠杆菌中的表达[J].重庆医科大学学报,2004(02):124-126.

HE Yujuan, et al. The expression of open reading frame of unknown KH gene from K562 cells in E.coli. [J]. Journal of Chongqing Medical University (2):124-126(2004)
[1]何於娟,欧一衡,张彦,刘小珊,马凌娣,涂植光,蒋纪恺.K562细胞未知基因KH的开放阅读框架在大肠杆菌中的表达[J].重庆医科大学学报,2004(02):124-126. DOI： 10.13406/j.cnki.cyxb.2004.02.002.

HE Yujuan, et al. The expression of open reading frame of unknown KH gene from K562 cells in E.coli. [J]. Journal of Chongqing Medical University (2):124-126(2004) DOI： 10.13406/j.cnki.cyxb.2004.02.002.

摘要

目的 :构建K5 6 2细胞未知基因KH的开放阅读框架（openreadingflame

ORF）的原核表达体系。方法 :采用DNA重组技术构建KH基因片段的开放阅读框架（KH -ORF）的表达载体pCI-neo-KH -ORF

热休克法转化大肠杆菌BL2 1（DE3）plysS

IPTG诱导后

SDS -PAGE鉴定目的蛋白的表达。结果 :经IPTG诱导 0 .5h后KH -ORF在BL2 1（DE3）plysS中获得表达

表达产物以包涵体形式存在

分子产量约 2 4kD。结论 :成功构建了KH -ORF的原核表达体系

为进一步研究其功能奠定了基础。

Abstract

Objective:To construct the prokaryotic expression system of open reading frame of KH gene (KH-ORF)

an unknown gene in K562 cells.Methods:The 225bp KH-ORF cDNA was cleaved from plasmid pGEM-T-easy-KH-ORF with EcoRI and SalI and subcloned into the pCI-neo Mammalian Expression Vector cutted with the same restriction endonucleases.The recombinant expression plasmid was transfected into BL21 (DE3) plysS by hot-shock assay.Induced by IPTG

the recombinant protein product was tested by SDS-PAGE.Results:24kD KH-ORF product was obtained as inclusion bodies in BL21 (DE3) plysS after the expression vector had been induced by IPTG for 30 minutes Conclusion :The construction of prokaryotic expression system of KH- ORF is successful.

关键词

K562细胞KH基因开放阅读框架原核表达

Keywords

K562 cellsKH geneOpen reading frameprokaryotic expression

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重庆医科大学临床学院消化内科

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