Secreted expression of Dengue virus type Ⅰ envelope glycoprotein in 293T cells
10.3760/cma.j.issn.1003-9279.2009.06.005
- VernacularTitle:登革Ⅰ型病毒E蛋白在293T细胞中的分泌表达
- 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 HANG
;
De-Xin LI
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Dengue virus;
Gene;
PrM/E;
Expression sequence tags
- From:
Chinese Journal of Experimental and Clinical Virology
2009;23(6):415-417
- CountryChina
- Language:Chinese
-
Abstract:
Objective To expression prM/E gene of dengue vires type I in mammalia cells.Methods The 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 poDNAS/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.Results In the cytoplasm of 293T cells transfected with all the recombinant plasmids DNA,the expressed producm 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 supematants transfected with the modified recombinant plasmids DNA.Conclusion The 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.
