Expression and bioassay of rubella virus E1-374 glycoprotein in yeast cells.
- Author:
Zhen-Mei LI
1
;
Hong-Ling WEN
1
;
Bin LIN
1
;
Cheng-Xi SUN
1
;
Fu-Lu CHU
1
;
Xiao-Jing YUAN
1
;
Yan-Yan SONG
1
;
Hong-Zhi XU
1
;
Zhi-Yu WANG
1
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression; Humans; Mice; Mice, Inbred BALB C; Pichia; genetics; metabolism; Recombinant Proteins; genetics; immunology; Rubella virus; genetics; immunology; Viral Envelope Proteins; genetics; immunology
- From: Chinese Journal of Experimental and Clinical Virology 2013;27(4):295-297
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo express the rubella virus E1-374 glycoprotein in Pichia pastoris and study the immunogenecity of the recombinant protein.
METHODSThe cDNA of protein E1-374 was cloned into the expression vector pGAPZalphaA and transformed into Pichia pastoris GS115 cells by electrotransfection. The expressed protein was confirmed by indirect immunofluorescence and demonstrated immunoreactivity by Western Blot. Rubella virus IgG antibody was assayed with ELISA after mice were inmmunized by E1-374 glycoprotein.
RESULTSSDS-PAGE analysis and Western Blot analysis of E1-374 protein revealed this protein to be 46.89 x 10(3). Antiserum (1:100) and E1-374 (5.5 microg/ml) was chosen for ELISA optimization. The intra-assay coefficient of variation for the ELISA was 0.36%-12.45%.
CONCLUSIONProtein E1-374 was highly expressed in Pichia pastoris cells, and it was a good choice to prepare rubella virus recombinant protein vaccines.
