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Microbiology and Biotechnology Letters

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Microbiol. Biotechnol. Lett. 1986; 14(6): 455-460

재조합 효모 세포내에서의 간염백신 생산

The Production of HBsAg in the Recombinant Yeast Cells

Cha-Yong Park and Hei-Chan Lee

Department of Chemical Technology College of Engineering Seoul National University, 1Department of Chemical Technology College of Engineering Seoul National University

Abstract

Dane particle was prepared from the plasma of chronic HBsAg carrier with high levels of HBsAg activity. DNA extracted front Dane particle core after a DNA polymerase reaction with $alpha$-($^{32}$P) dNTP, was identified as HBV DNA by liquid scintillation counter and agarose gel electrophoresis-G.M. counting. To produce Hepatitis B surface antigen for use as a vaccine against Hepatitis B virus infection, yeast strains harboring recombinant plasmid with Apase promoter was used. Recombinant plasmid was construced from pHBV 130 and pAN 82, transformed into E coli, and then transferred into yeast strains. HBsAg was produced by derepression in Burkholder minimal medium with controlled inorganic phosphate concentration. The kinetics of HBsAg production was also investigated. Total HBsAg activity increased rapidly between 3 and 6 hours after transfer to phosphate-free medium and reached a maximum at around 9th hour. The transfer into phosphate-free medium after 6 hours in high phosphate cell growth medium gave maximum activity.

Keywords: HBsAg, Apase promoter, recombinant plasmid, bioreactor operation, yeast

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