Stable cell line for secretion of replication-defective hepatitis B virus vector expressing blasticidin resistant gene

VernacularTitle:稳定分泌重组HBV载体颗粒细胞系的构建
Author: Jin-Xia LIU ¹ ; Dian-Xing SUN ; Zhi-Chen CAO
Author Information

1. 河北医科大学
Keywords: Hepatitis B virus; Cell Line; Magnaporthe
From: Chinese Journal of Experimental and Clinical Virology 2009;23(4):316-318
CountryChina
Language:Chinese
Abstract: Objective To construct a stable cell line with permanent secretion of recombinant hepatitis B virus(HBV) vector, which express blasticidin resistant gene. Methods Replication-defective HBV vector, pCH-BsdR, which express blasticidin resistance gnne was constructed by deleting the HBV genes and inserting the blasticidin resistance gene into the S region. The CA18-resistant, the packaging signal deleted HBV helper plasmid, pcDNA3.1-CH3142, and the HBV vector pCH-BsdR were cotransfectcd into HepG2 cells. Cell clones were selected by the adding of both blasticidin and G418, then serial detection were done.Results After 36 cell clones were picked and expanded. Three cell clones were defined as the best. Quantity of their HBV DNA were 4.1 × 106, 3.6×106 and 1.2× 106 copies/ml, respectively. Enveloped recombinant, but not wild type HBV were confirmed in the culture medium. Conclusions The stable cell lines can realize large preparation of recombinant HBV virions. This will contribute to the use of HBV vector for gene therapy and HBV suseeptihle cell lines screening.