Stable cell line for secretion of replication-defective hepatitis B virus vector expressing blasticidin resistant gene / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
; (6): 316-318, 2009.
Article
in Zh
| WPRIM
| ID: wpr-325555
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct a stable cell line with permanent secretion of recombinant hepatitis B virus (HBV) vector, which express blasticidin resistant gene.</p><p><b>METHODS</b>Replication-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.</p><p><b>RESULTS</b>After 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.</p><p><b>CONCLUSIONS</b>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 susceptible cell lines screening.</p>
Full text:
1
Index:
WPRIM
Main subject:
Pharmacology
/
Physiology
/
Virion
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Virology
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Virus Replication
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Drug Resistance
/
Transfection
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Genetic Engineering
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Gene Expression
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Cell Line
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2009
Type:
Article