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KMID : 0366119860140060455
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Korean Journal of Applied Microbiology & Bioengineering
1986 Volume.14 No. 6 p.455 ~ p.460
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The Production of HBsAg in the Recombinant Yeast Cells
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Choi Cha-Yong
Lee Hei-Chan
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Abstract
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Dane particle was prepared from the plasma of chronic HBsAg carrier with high levels of HBsAg activity. DNA extracted from Dane particle core, after a DNA polymerase reaction with Ct-(:n2p) 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 pAM 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 in-creased rapidly between 3 and 6 hours after transfer to phosphate-free medium and reached a maximum at around 9th hour. The transfer into phosphale-free medium after 6 hours in high phosphate cell growth medium gave maximum activity.
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KEYWORD
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HBsAg, Apase promoter, recombinant plasmid, bioreactor operation, yeast
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