Selection and characterization of packaging cell lines for XJ-160 virus.
OBJECTIVE
XJ-160 virus is a mosquito-derived Sindbis-like virus isolated in China. Based on its infectious clone (pBR-XJ-160) we have developed an RNA-based vector system. In this work, we constructed packaging cell lines (PCLs) for XJ-160 virus.


METHODS
Firstly, XJ-160 virus structural protein expression cassette pcE or pICH was constructed by cloning the gene of glycoprotein E3E26KE1 or capsid protein of XJ-160 virus into pcDNA3.1(+) or pIRES, respectively. Then the PCLs (BHK-21(E+Capsid) cells) for XJ-160 virus were obtained by two selections with G418 and hygromycin.


RESULTS
The results indicate that BHK-21(E+Capsid) cells, stably expressing E3E26KE1 protein and capsid protein of XJ-160 virus, not only highly increased packaging efficiency of the vector from XJ-160 virus, but also provided packaging function for the vector from Semliki Forest virus.


CONCLUSIONS
These results suggest potential utility of the PCLs of XJ-160 virus for large-scale vector production and facilitating broad alphavirus applications. Also, the construction of PCLs for XJ-160 virus lays a basis for developing alphavirus-derived vector systems.
State Key Laboratory for Infectious Disease Prevention and Control, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 100 Yingxin Street, Beijing, China.
Wu-yang
WY
Zhu
Guo-dong
GD
Liang
D000818:Animals; D002460:Cell Line; D002681:China; D006224:Cricetinae; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D012672:Semliki forest virus / Q000502:physiology*; D012845:Sindbis Virus / Q000235:genetics*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D019065:Virus Assembly
D014764:Viral Proteins