Construction and identification of prokaryotic expression vector of human cystatin C

WANG jian

doi:10.13406/j.cnki.cyxb.2008.03.007

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Construction and identification of prokaryotic expression vector of human cystatin C

更新时间：2024-01-17

- Construction and identification of prokaryotic expression vector of human cystatin C
- Journal of Chongqing Medical University Issue 3, Pages: 331-333(2008)
- 作者机构：
  
  1. 重庆医科大学医学检验系临床检验诊断学省部共建教育部重点实验室
  2. 重庆医科大学医学检验系临床检验诊断学省部共建教育部重点实验室,重庆,400016
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2008.03.007
  CLC： R346
- Published：2008，
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[1]王箭,阳强,陈婷梅,尹一兵.Cys C的原核表达载体构建和鉴定[J].重庆医科大学学报,2008(03):331-333.

WANG jian. Construction and identification of prokaryotic expression vector of human cystatin C. [J]. Journal of Chongqing Medical University (3):331-333(2008)
[1]王箭,阳强,陈婷梅,尹一兵.Cys C的原核表达载体构建和鉴定[J].重庆医科大学学报,2008(03):331-333. DOI： 10.13406/j.cnki.cyxb.2008.03.007.

WANG jian. Construction and identification of prokaryotic expression vector of human cystatin C. [J]. Journal of Chongqing Medical University (3):331-333(2008) DOI： 10.13406/j.cnki.cyxb.2008.03.007.

摘要

目的:构建人胱抑素C（Cys C）的原核表达载体

为进一步表达纯化Cys C重组蛋白奠定基础。方法:从HL-60细胞中提取总RNA

RT-PCR扩增Cys C基因

再将目的基因克隆至PET32a（+）表达载体中

构建人Cys C原核表达质粒PET32a（+） /Cys c;再将此重组子进行双酶切电泳鉴定和测序鉴定。结果:酶切电泳鉴定结果显示Cys C基因克隆入载体中

测序结果证实克隆的基因序列与GeneBank中的Cys C序列相符。结论:获得了人Cys C原核表达质粒PET32a（+）/Cys C

为进一步表达和纯化Cys C重组蛋白奠定了工作基础。

Abstract

Objective:To Construct a prokaryotic expression vector of cystatin C(Cys C)to provide a basis of purify Cys C protein produced by the expression system.Methods:Total RNA was isolated from HL-60 cells

and human Cys C gene was amplified with RT-PCR.The cDNA fragment was cloned into pET32a(+)vector

which was confirmed with sequencing and electrophoresis.Results:The result of electrophoresis showed that Cys C had been cloned into pET32a(+)vector

and the result of.DNA sequence analysis also showed the cloned Cys C gene sequence was completely correspondent to GeneBank data.Conclusion:The prokaryotic expression vector of cystatin C(Cys C)was obtained

providing a basis of purification of Cys C protein produced by the expression system.

关键词

胱抑素C原核表达系统

Keywords

cystatin CProkaryotic expression system

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Related Author

NIE Wen-rong

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Department of Pediatrics,the People’s Hospital of Qiyang County in Hunan Province

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