Selection and characterization of packaging cell lines for XJ-160 virus

Abstract

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-21E+Capsid cells) for XJ-160 virus were obtained by two selections with G418 and hygromycin. Results: The results indicate that BHK-21E+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. Copyright © 2009 S. Karger AG, Basel.