[Study on E protein epitopes and primary identification of main yellow virus].
OBJECTIVE
To analysis the E protein epitopes of dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus and to distinguish the shared or specific epitopes among them.


METHODS
Bioinformatic software DNAStar was used to analyze the hydrophilicity, flexibility, surface probability and antigenicity of dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus E protein amino acid sequences. The influence of secondary structure was also considered. Based on the bio-informatic analysis of E protein epitopes, 6 specific epitopes were amplified and inserted into prokaryotic expression vector pMAL-c2x. The vectors was then transferred into E. coli BL21 (DE3) and Rosetta (DE3). Isopropyl-beta-D-thiogalactoside (IPTG) was used to induce the expression of gene segments and SDS-PAGE were used identify the expression proteins. The antigenicity was tested, using Western blot.


RESULTS
15 shared epitopes and 47 specific epitopes were forecasted by bioinformatic analysis, and 6 specific epitopes from dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus E protein were expressed in E. coli successfully. Two specific antigenic determinant from dengue virus type 1 and dengue virus type 2 were confirmed using Western blot, while the others epitopes shown no antigenic reaction property.


CONCLUSIONS
Two specific antigenic determinant were confirmed, under Western blot.
Instrumental Analysis & Research Center of South China Agricultural University, Guangzhou 510642, China.
Xiao-li
XL
Xu
Jian-jun
JJ
Yang
Rui-wen
RW
Ren
Jian-wei
JW
Liu
Si-bei
SB
Ma
Zhi-jun
ZJ
Bai
Mei-yu
MY
Fang
D000956:Antigens, Viral / Q000235:genetics; D015153:Blotting, Western; D003716:Dengue Virus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D004664:Encephalitis Virus, Japanese / Q000235:genetics* / Q000276:immunology; D000939:Epitopes / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology; D015005:Yellow fever virus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D000956:Antigens, Viral; D000939:Epitopes; D014759:Viral Envelope Proteins