2002•Academic Journal of Second Military Medical UniversityRequires access

Establishment and identification of P815 cell line stably expressing HBsAg

Feng Zhou

Abstract

Objective: To screen for and establish P815 cell line stably expressing HBsAg (adr subtype) which is com-mon in Asian people, providing the cell infection model for evalution of the hepatitis B DNA vaccine effect in virto. Methods:S gene of hepatitis B virus was amplified and cloned into pcDNA3. The recombinant plasmid was transfected P815 by lipo-some and screened by G418. Results: Recombinant plasmid pcDNA3-HBsAg(S) coding hepatitis B (adr subtype) surface pro-tein S was constructed. HBsAg was detected both in the trans fected cell and its culture medium. It was confirmed by PCRthat HBsAg(S) gene was integrated into the chromosome of P8l5 stably. Conclusion: P815 cell line stably expressing HBsAgis obtained, providing bases for detecting the CTL reaction stimulated by hepatitis B DNA vaccine.

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Key concepts: HBsAg, Transfection, Virology, Plasmid, Recombinant DNA, Molecular biology, Cell culture, CTL*