Construction of eukaryotic expression vector of open reading frame of unknown KH gene and its effect on proliferation of K562 cell

HE Yujuan; et al

doi:10.13406/j.cnki.cyxb.2004.05.001

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Construction of eukaryotic expression vector of open reading frame of unknown KH gene and its effect on proliferation of K562 cell

更新时间：2024-01-17

- Construction of eukaryotic expression vector of open reading frame of unknown KH gene and its effect on proliferation of K562 cell
- Journal of Chongqing Medical University Issue 5, Pages: 559-562(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. 重庆医科大学临床学院血液科
  3. 重庆市永川市人民医院
  4. 重庆医科大学基础医学院临床生化教研室,重庆,400016
  5. 永川,重庆,402160
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2004.05.001
  CLC： Q781
- Published：2004，
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[1]何於娟,张彦,刘小珊,马凌娣,欧一衡,蒋纪恺.未知基因KH开放阅读框架真核表达载体的构建及其对K562细胞增殖的影响[J].重庆医科大学学报,2004(05):559-562.

HE Yujuan, et al. Construction of eukaryotic expression vector of open reading frame of unknown KH gene and its effect on proliferation of K562 cell. [J]. Journal of Chongqing Medical University (5):559-562(2004)
[1]何於娟,张彦,刘小珊,马凌娣,欧一衡,蒋纪恺.未知基因KH开放阅读框架真核表达载体的构建及其对K562细胞增殖的影响[J].重庆医科大学学报,2004(05):559-562. DOI： 10.13406/j.cnki.cyxb.2004.05.001.

HE Yujuan, et al. Construction of eukaryotic expression vector of open reading frame of unknown KH gene and its effect on proliferation of K562 cell. [J]. Journal of Chongqing Medical University (5):559-562(2004) DOI： 10.13406/j.cnki.cyxb.2004.05.001.

摘要

目的 :构建未知基因KH的开放阅读框架（openreadingframe

ORF）的真核表达体系

初步探讨未知基因KH的功能。方法 :采用DNA重组技术将KH基因片段的开放阅读框架（KH -ORF）亚克隆于表达载体 pCl-neoMammalianExpres sionVector

重组为pCI-neo -KH -ORF真核表达体系

脂质体转染K5 6 2细胞

RT -PCR检测目的基因KH的表达。采用体外培养技术

通过 pCI -neo -KH -ORF质粒转染K5 6 2细胞前后的生长曲线和乳酸脱氢酶活性测定观察KH -ORF对K5 6 2细胞增殖速度的影响。结果 :KH -ORF在K5 6 2细胞中获得表达

对细胞增殖速度有明显影响。结论 :KH -ORF表达产物可能是一个与K5 6 2细胞增殖有关的功能蛋白质

KH基因可能是一个具有生物学功能的白血病未知基因

Abstract

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

an unknown gene obtained from K562 cells

and investigate its function.Methods:The KH-ORF cDNA was cleaved from plasmid expression vector cut with the same restriction endonucleases.pCI-KH-ORF

the pGEM-T-easy-KH-ORF with EcoRI and SalI and subcloned into the pCl-neo mammalian recombinant expression plasmid

was transfected into K562 cells with liposome and screened by G418.KH-ORF expression was tested by RT-PCR.The effect of expression product on cell proliferation was tested by growth-curve and LDH detection.Results:KH-ORF was expressed in K562 cells.The effects of increased proliferation were significant.Conclusion:The product of KH-ORF may be a functional protein that is related to the proliferation of K562 cells

and KH gene may be a functional unknown leukemia gene.

关键词

KH基因开放阅读框架真核表达K562细胞增殖

Keywords

KH geneOpen reading frameEukaryotic expressionK562 cellsproliferation

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Related Author

HE Yujuan

et al

张彦

刘小珊

马凌娣

涂植光

蒋纪恺

Chen Tingmei

Related Institution

Department of Gastrointestinal Surgery，The Third Affiliated Hospital of Zunyi Medical University/ The First People’s Hospital of Zunyi

Department of Obstetrics and Gynecology,Yongchuan Hospital Affiliated to Chongqing Medical University

Department of Obstetrics and Gynecology,The First Affiliated Hospital of Chongqing Medical University

Central Laboratory of Yongchuan Hospital Affiliated to Chongqing Medical University

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