[Identification and construction of replicon vectors of Japanese encephalitis virus (strain SA14-14-2)].
OBJECTIVE
In order to lay the groundwork for studying the novel vaccine Identified.


METHODS
(1) Two replicons were constructed. One's prM/E gene was deleted completely (Full AprM/E Replicon), the other's prM/E gene was deleted partially (213 bp of C terminal of E gene was reserved; Partial delta prM/E Replicon), and the deleted parts was replaced as the MCS. (2) Replicons RNA were which will use the JEV as the vector, replicon vectors of JEV was constructed and transfected into BHK-21 cell. After 24, 48, 72, 96 h, method of real-time PCR was used to identify Replicons' replication ability. (3) YFP gene was inserted into the MCS of those two replicons. Their RNA was transfected into BHK-21 cell. Expression of YFP was tested by the fluorescence microscopy and flow cytometer.


RESULTS
(1) After the two replicons RNA were transfected into BHK-21 cell, as time went by, the quantity of RNA increased. (2) After RNA of the replicons with YFP were transfected into BHK-21 cell, increasing trend of fluorescent signal and rate of YFP positive cell was observed and tested.


CONCLUSIONS
Full delta prM/E Replicon and Partial delta prM/E Replicon have the ability to duplicate itself and express the foreign protein.
Wei
Yan
Y
State Key Laboratory for Infectious Disease Prevention and Control, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing.
Li
Chuan
C
Lu
Peng
P
Yu
Jian-shi
JS
Li
Jian-dong
JD
Liu
Qin-zhi
QZ
Zhang
Quan-fu
QF
Li
De-xin
DX
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004261:DNA Replication; D004664:Encephalitis Virus, Japanese / Q000235:genetics* / Q000378:metabolism; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D012093:Replicon