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KMID : 0545119910010010031
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Journal of Microbiology and Biotechnology
1991 Volume.1 No. 1 p.31 ~ p.36
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Secretion of Bovine β-Casein by Saccharomyces cerevisiae
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Chung, Kun Sub
Rafael, F. R./Oh, Sang Suk/Richardson, T.
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Abstract
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Yeast expression plasmids containing an appropriate leader sequence and bovine β-casein cDNA were constructed to produce β-casein for the study of its functional characteristics. Two kinds of expression systems for β-casein were constructed using pCGY1444 as a precursor plasmid. This plasmid is a yeast-E. coli shuttle vector which contains the chelatin promoter. The plasmid pISB202 contains the invertase leader sequence and β-casein gene. The plasmid pDEB303 contains the original bovine β-casein leader sequence gene. These two plasmids were introduced into S. cerevisiae AB116 which is a strain deficient in the major yeast proteinases. Each clone was grown in minimal media for 24h before induction by CuSO_4. The cells were thus grown under expression conditions. Both strains harbouring pISB202 and pDEB303 expressed bovine β-casein. The β-casein was detected using immunochemical staining after western blot. Secretion of β-casein was detected in the culture broth. The estimated amount of secreted β-casein was approximately 50㎍/ℓ.
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