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Published on: June 19, 2013
Selection and characterization of packaging cell lines for XJ-160 virus
1State 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.
Objectives:
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.
Conclusion:
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.
