[Construction and characterization of reverse genetic system for H9N2 avian influenza].
OBJECTIVE
To provide a technology platform for vaccine development as well as the research on transmission and pathogenesis, the reverse genetic system for H9N2 avian influenza virus was established.


METHODS
Eight full-length cDNAs of avian influenza virus A/Guangzhou/333/99 (H9N2) were amplified by RT-PCR and separately cloned into the transcription/expression vector, pCI-polI. The 8 plasmids DNA was cotransfected into 293T cell, the cell supernatant was collected and inoculated into embryonated eggs, the rescued virus from the allantoic fluid was identified by hemagglutinination assay.


RESULTS
The avian influenza H9N2 virus was successfully rescued by 8 plasmids co-transfection in 293T cells. The hemagglutinination titer of the rescued virus is up to 2(9)/50 microl and its growth curve remained relatively as to the wild-type virus.


CONCLUSIONS
The reverse genetic for avian influenza H9N2 subtype virus has been established successfully.
Liu
Li-Qi
LQ
Chinese Influenza Center, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Dong
Jie
J
Bo
Hong
H
Wen
Le-Ying
LY
Xin
Li
L
Zhang
Ye
Y
Li
Zi
Z
Dong
Li-Bo
LB
Wang
Min
M
Guo
Yuan-Ji
YJ
Shu
Yue-Long
YL
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D005260:Female; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007223:Infant; D053127:Influenza A Virus, H9N2 Subtype / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D007251:Influenza, Human / Q000821:virology; D010957:Plasmids / Q000235:genetics