Cloning of envelope glycoprotein K8.1﹑K8.1A and gL gene of KSHV and eukaryotic expression
DOI:
CSTR:
Author:
Affiliation:

Clc Number:

Fund Project:

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

    Objective:To clone envelope glycoprotein K8.1,K8.1A and gL gene of Kaposi’s sarcoma-associated herpesvirus(KSHV)and express them in 293T cells. Methods:K8.1 and gL genes were amplified by PCR with the total DNA of BCBL-1 cells as template. K8.1A genes were amplified by RT-PCR with the total RNA of BCBL-1 cells as template. The PCR fragments were cloned into pcDNA3.1(+) vector to construct recombinant eukaryotic expression plasmids. 293T cells were transfected with the recombinant plasmids. Western blot was performed to evaluate the expressions of K8.1 and K8.1A in 293T cells. RT-PCR was performed to evaluate the transcription of gL in 293T cells. Results:The sequence of K8.1,K8.1A and gL was 100% homology with K8.1,K8.1A and gL gene of KSHV previously registered in GenBank. An interesting band about 35 ku was visible in the result of Western blot,which was consistent with expected size of recombinate protein of K8.1 and K8.1A expressed in 293T. An interesting band about 500 bp was visible in the result of RT-PCR. Conclusion:Glycoprotein K8.1,K8.1A and gL of KSHV were correctly expressed in 293T cells.

    Reference
    Related
    Cited by
Get Citation

徐华国,卢春,程林,曾怡,秦娣,吕志刚,陈秀英. KSHV包膜糖蛋白编码基因K8.1、K8.1A和gL的克隆及真核表达[J].南京医科大学学报(自然科学版英文版),2007,(5):502-506.

Copy
Share
Article Metrics
  • Abstract:
  • PDF:
  • HTML:
  • Cited by:
History
  • Received:December 30,2006
  • Revised:
  • Adopted:
  • Online:
  • Published:
Article QR Code