DNA for expression of aprotinin in methylotrophic yeast cells

Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Fungi

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4353201, 530300, 536 235, C07K 710, C12N 119, C12N 1515, C12P 2102

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active

052583023

ABSTRACT:
Biologically active aprotinin (APR) molecules, naturally occurring, relatively short, single chain polypeptides, are prepared by growing methylotrophic yeast transformants containing in their genome at least one copy of a DNA sequence operably encoding APR, in operational association with a DNA sequence encoding the S. cerevisiae alpha mating factor pre-pro sequence (including a processing sequence selected from lys-arg-(glu-ala).sub.x, wherein x is an integer falling in the range of 0-3), both under the regulation of a promoter region of a gene of a methylotrophic yeast, under conditions allowing expression of said DNA sequences, and secretion of APR molecules into the culture medium.
Also disclosed are novel DNA fragments and novel recombinant yeast strains which are useful in the practice of the present invention.

REFERENCES:
patent: 4837148 (1989-06-01), Cregg
patent: 4855231 (1989-08-01), Stroman et al.
patent: 4882279 (1989-11-01), Cregg
patent: 4929555 (1990-05-01), Cregg et al.
Norris et al. "Aprotinin and Aprotinin Analogues Expressed in Yeast" Biolcal Chemistry Hoppe-Seyler 371 (suppl):37-42 (Jun. 1989).
Kingsman et al. "Heterologous Gene Expression in S. cerevisiae" Biotechnology and Genetic Engineering Reviews 3:377-416 (Sep. 1985).

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