[Molecular characteristics of the full-length genome of two new Japanese encephalitis virus isolated from mosquitoes in Hubei province].
OBJECTIVE
To sequence and analyze the complete genome of two new Japanese encephalitis virus (JEV) strains isolated from mosquitoes collected in Hubei province in 2008, and to understand the molecular biological characteristics of JEV in this area.


METHODS
RT-PCR was used to amplify the fragments of HBZG08-09 strain and HBZG08-55 strain with 16 pairs overlapping primers after they had been recovered and identified, then the full-length genome was obtained by sequencing and splicing. Biological sequence alignment, nucleotide and amino acid sequence analysis, phylogenetic analysis and analysis of amino acid differences were performed by the software of Clustal X (1.83), MegAlign, Mega (4.0) and Genedoc (3.2).


RESULTS
The genome of two new strains were both 10 965 nucleotides in length with a single open reading frame from 96 to 10 392 coding for a 3432 amino acid poly-protein, the homology of nucleotide and amino acid sequence between two isolates were 98.2% and 99.7% respectively. Further study showed that the new strains were both belonging to genotype I. Two new strains were most closely related to isolates obtained from Henan and Zhejiang province in recent years. Compared with the live attenuated vaccine strain SA-14-14-2 in China, HBZG08-09 strain had 82 amino acid divergence; HBZG08-55 had 84 amino acid divergences. But the amino acid difference occurred in sites were not the key ones affecting the toxicity or antigenic of JEV.


CONCLUSIONS
Two new JEV isolates were both belonging to genotype I, and the key sites of amino acid were not changed.
Jiang
Hong-Yue
HY
Department of Virology, School of Public Health, Shandong University, Jinan 250012, China.
Li
Ming-Hua
MH
Yang
Xiao-Bing
XB
Li
Pei-Long
PL
Zhang
Jing
J
Meng
Feng-Xia
FX
Guo
Yu-Hong
YH
Mei
Li-Zhi
LZ
Fu
Shi-Hong
SH
He
Yuan-Yuan
YY
Wang
Huan-Yu
HY
Wang
Zi-Jun
ZJ
Yang
Wei-Zhong
WZ
Wang
Zhi-Yu
ZY
Liang
Guo-Dong
GD
D000818:Animals; D002681:China; D009033:Culicidae / Q000821:virology*; D004664:Encephalitis Virus, Japanese / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016679:Genome, Viral; D005838:Genotype; D008969:Molecular Sequence Data; D010802:Phylogeny; D012367:RNA, Viral; D016415:Sequence Alignment
D012367:RNA, Viral