Detection of the expression of alpha3-integrin on hantavirus permissive cells.
- Author:
Jingfang DONG
1
;
Yuqing JIANG
;
Jianwei WANG
;
Yanxiang XIE
;
Changshou HANG
;
Tao HONG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cercopithecus aethiops; Cloning, Molecular; Gene Expression; Hantavirus; Integrin beta3; biosynthesis; genetics; immunology; Rabbits; Receptors, Virus; Vero Cells; metabolism
- From: Chinese Journal of Experimental and Clinical Virology 2002;16(1):40-43
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUNDTo express and purify human alpha3-integrin to serve as the antigen to prepare its antibody and to separate the Vero cell clones without expression of alpha3-integrin.
METHODSThe human alpha3-integrin gene was amplified by using RT-PCR, then subcloned into a pQE30 expression vector and expressed in E. coli. The gene expression was confirmed by Western blot assay. Rabbit was inoculated with purified antigen to stimulate the antibody generation. The target Vero cells were separated by negative selection using antibody plus complement mediated cytolysis. The separated cell clones were confirmed by immunofluorescence and Western blot assay.
RESULTSThe alpha3- integrin gene was cloned and expressed effetively, Western blot assay revealed that the expressed protein held good immune reactivity. High titer antibody was generated. However the expression of alpha3-integrin was not detected on Vero, VeroE6, Hep-2, 2BS and 293 cells.
CONCLUSIONSThe results of the study suggested that hantavirus has other receptors on Vero cells beside alpha 3-integrin.
