Secreted expression of dengue virus type I envelope glycoprotein in 293T cells.
- Author:
Fang MIAO
1
;
Chuan LI
;
Shuo ZHANG
;
Xiao-fang WANG
;
Jian-dong LI
;
Quan-fu ZHANG
;
Qin-zhi LIU
;
Yan WEI
;
Xiao-tong HANG
;
Mi-fang LIANG
;
De-xin LI
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; Dengue; virology; Dengue Virus; genetics; metabolism; Gene Expression; Glycoproteins; genetics; metabolism; Humans; Protein Transport; Recombinant Fusion Proteins; genetics; metabolism; Viral Envelope Proteins; genetics; metabolism
- From: Chinese Journal of Experimental and Clinical Virology 2009;23(6):415-417
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo expression prM/E gene of dengue virus type I in mammalia cells.
METHODSThe full-length prM/E gene of dengue virus type I strain GZ01/95 was amplified by RT-PCR, the signal peptide preceding the prM gene was added or the carboxyl-terminal 20% of DEN-1 E was replaced with the corresponding JE sequence in the meanwhile, and three of the constructions were cloned into the pcDNA5/FRT.Then they were transfected into 293T cells by lipofectamine respectively. The expression of recombinant proteins were identified by indirect immuno-fluorescence assay(IFA) as well as Western blot.
RESULTSIn the cytoplasm of 293T cells transfected with all the recombinant plasmids DNA, the expressed products for gene of dengue virus type I were confirmed by IFA. The secreted expression products for gene of dengue virus type I specific protein bands were confirmed by Western blot only existing in the cell supernatants transfected with the modified recombinant plasmids DNA.
CONCLUSIONThe prM/E protein of dengue virus type 1 were expressed in 293T cells transfected with all the three recombinant plasmids DNA. The prM/E protein was obtained secretion after transfecting the modified recombinant plasmids adding a signal peptide preceding the prM gene or replacing the carboxyl-terminal 20% of E with the corresponding JE sequence.
