Stable cell line for secretion of replication-defective hepatitis B virus vector expressing blasticidin resistant gene.
- Author:
Jin-Xia LIU
1
;
Dian-Xing SUN
;
Zhi-Chen CAO
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; drug effects; virology; Clone Cells; Drug Resistance; Gene Expression; Genetic Engineering; Genetic Vectors; genetics; metabolism; Hep G2 Cells; Hepatitis B virus; genetics; physiology; Humans; Nucleosides; pharmacology; Transfection; Virion; genetics; physiology; Virus Assembly; Virus Replication
- From: Chinese Journal of Experimental and Clinical Virology 2009;23(4):316-318
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a stable cell line with permanent secretion of recombinant hepatitis B virus (HBV) vector, which express blasticidin resistant gene.
METHODSReplication-defective HBV vector, pCH-BsdR, which express blasticidin resistance gene was constructed by deleting the HBV genes and inserting the blasticidin resistance gene into the S region. The G418-resistant, the packaging signal deleted HBV helper plasmid, pcDNA3.1-CH3142, and the HBV vector pCH-BsdR were cotransfected into HepG2 cells. Cell clones were selected by the adding of both blasticidin and G418, then serial detection were done.
RESULTSAfter 36 cell clones were picked and expanded. Three cell clones were defined as the best. Quantity of their HBV DNA were 4.1 x 10(6), 3.6 x 10(6) and 1.2 x 10(6) copies/ml, respectively. Enveloped recombinant, but not wild type HBV were confirmed in the culture medium.
CONCLUSIONSThe 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 susceptible cell lines screening.
