Construction of the eukaryote expression vector carrying encoding gene of human interleukin-1 receptor antagonist and its transient expression in human gingival fibroblast cells
DOI:
CSTR:
Author:
Affiliation:

Clc Number:

Fund Project:

  • Article
  • |
  • Figures
  • |
  • Metrics
  • |
  • Reference
  • |
  • Related
  • |
  • Cited by
  • |
  • Materials
  • |
  • Comments
    Abstract:

    Objective:To construct the recombinant pEGFP-N1/hIL-1ra and test its transient expression in human gingival fibroblast cells, this may be useful of the treatment of periodontitis. Methods:Total RNA was extracted from human mammary cancer tissue. RT-PCR was performed to amplify the hIL-1ra encoding gene. Then PCR product was purified and cloned into pMD18-T. After DNA sequencing,both of the recombinant vector pMD18-T/hIL-1ra and plasmid pEGFP-N1 were digested with HindⅢ and BamHⅠ respectively. IL-1ra gene fragment was ligated into plasmid pEGFP-N1. The recombinant plasmid DNA was transformed into E.coli competent cells TOP 10 and positive clones were selected and tested. After being transfected by pEGFP-N1/hIL-1ra,the transient expression of IL-1ra gene in human gingival fibroblast(HGF) cells were detected by fluorescence microscope and RT-PCR. Results:hIL-1ra encoding gene fragment was 531 bp and was inserted into the eukaryotic expression vector pEGFP-N1/hIL-1ra correctly. And HGF cells,which were identified by both fluorescence microscope and RT-PCR,had a transient expression of IL-1ra after transfection. Conclusion:The new recombinant expression vector pEGFP-N1/hIL-1ra was constructed successfully and the HGF cells which could express IL-1ra transiently were also successfully made.

    Reference
    Related
    Cited by
Get Citation

苏 毅,何雅丽,王子露,李建民,徐 艳. pEGFP-N1/hIL-1ra重组真核表达载体的构建及其在人牙龈成纤维细胞中的瞬时表达[J].南京医科大学学报(自然科学版英文版),2008,28(12):1533-.

Copy
Share
Article Metrics
  • Abstract:
  • PDF:
  • HTML:
  • Cited by:
History
  • Received:July 03,2008
  • Revised:
  • Adopted:
  • Online:
  • Published:
Article QR Code