J4 ›› 2009, Vol. 35 ›› Issue (5): 898-901.

• 基础研究 • 上一篇    下一篇

人纤溶酶真核表达重组质粒的构建

吴扬1|李玉琴2|王智昊1|臧秀贤1   

  1. 1.吉林大学第一医院急诊科|吉林 长春 130021;2.吉林大学第一医院胃肠内科|吉林 长春 130021
  • 收稿日期:2009-03-20 出版日期:2009-09-28 发布日期:2009-09-28
  • 通讯作者: 李玉琴 E-mail:liyuqin401@hotmail.com
  • 作者简介: 吴 扬(1960-)|女|吉林省长春市人|副教授|主要从事消化及心脑血管疾病的研究。
  • 基金资助:

    长春市科技计划项目资助课题(2006-2009)

Construction of eukaryotic expression recombinant plasmid of human fibrinolysin

WU Yang1,LI Yu-qin2,WANG Zhi-hao1,ZANG Xiu-xian1   

  1. 1.Department of Emergency,First Hospital,Jilin University,Changchun 130021,China;2.Department of Gastroenterology,First Hospital,Jilin University,Changchun 130021,China
  • Received:2009-03-20 Online:2009-09-28 Published:2009-09-28

摘要:

目的:构建人纤溶酶真核表达载体以用于抗栓、溶栓的研究。方法:人胎肝组织经PCR技术扩增人纤溶酶基因,定向克隆至真核表达质粒pC-DNA-3后测序。经测序鉴定正确后构建人纤溶酶真核表达重组质粒,利用PCR和酶切方法鉴定重组真核表达质粒的正确性。结果:目的基因 PCR扩增产物为1 740 bp,测序结果显示,目的基因PCR扩增产物与GenBank登记的序列完全相同。PCR和酶切鉴定结果显示,构建的重组真核表达质粒中含有正确编码的人纤溶
酶基因全长序列。结论:成功构建了含有人纤溶酶基因的真核表达重组质粒,并进行了鉴定。

关键词: 人纤溶酶;抗栓;真核表达载体;重组质粒

Abstract:

Abstract:Objective To construct the eukaryotic expression vector of human fibrinolysin for further  study on anti-thrombus and thrombolysis. Methods  Human fibrinolysin gene was amplified  from human fetal liver tissue with PCR,and was cloned into  eukaryotic expression plasmid pCI-neo and then sequenced.After the correct identification by sequencing,the eukaryotic expression recombinant plasmid of human fibrinolysin was constructed and identified with PCR and enzyme digestion.Results The PCR amplification product of fibrinolysin gene was 1 740 bp.The result of sequencing was the same as that registered in GenBank.The results of PCR and enzyme digestion showed that the recombinant eukaryotic expression plasmid had  correct codogenic gene fragment.Conclusion The eukaryotic expression recombinant plasmid of human fibrinolysin gene is successfully constructed and identified.

Key words:  human fibrinolysin;anti-thrombus;eukaryotic expression vector;recombinant plasmid

中图分类号: 

  • Q78