Yeast vector coding for heterologous gene fusions linked via KEX

Details Yeast vector coding for heterologous gene fusions linked via KEX Yeast vector coding for heterologous gene fusions linked via KEX

1995-02-23

1996-05-28

LeGuyader, John L.

Chemistry: molecular biology and microbiology

Vector, per se

435 691, 435 692, 435 697, 4351723, 435224, 43525421, 536 232, 536 234, 536 235, 536 242, C12N 1581, C12N 1552

Patent

active

055210937

ABSTRACT:
The present invention relates to an improved system for producing mature heterologous proteins, and especially hirudin, by means of yeasts comprising an expression vector containing a sequence coding for the heterologous protein. This improvement is characterized by amplification of the KEX2 gene of yeast, coding for the endoprotease yscF. The amplification is carried out either by integration of one or more copies of all or part of the KEX2 gene in the yeast genome, or by insertion of one or more copies of all or part of the KEX2 gene into the vector for expression of the heterologous protein.

REFERENCES:
Julius et al. Cell 37(3):1075-1090 (1984) abstract only.
Dmochowska et al. Cell 50(4):573-584 (1987) abstract only.
Loison et al. Biotechnology 6:72 (1988).
Bourbonnais et al. J. Biol Chem. 263:15342 (1988).
Steube et al. Yeast 4: (Sepc. Iss.) 5169 (1988).
Sleep et al. Yeast 4:(Sepc. Iss.) 5168 (1988).
Fuller et al, "The Yeast Pro-Hormone Cleaving Enzyme, KEX2", Abstruact X015, Journal of Cellular Biochemistry, Supplement 12B, Jan. 1988, p. 271.
Valls et al, "Protein Sorting in Yeast: The Localization Determinant of Yeast Vacuolar Carboxypeptidase Y Resides in the Propeptide", pp. 895-896.
Chemical Abstracts, vol. 98, No. 7, 14 Feb. 1983, p. 363, col. 2, No. 50144u, Jones et al, "Pleitropic mutations of S. cerevisiae which cause deficiency for proteinases . . . ".
Struhl, "Nucleotide sequence and transcriptional mapping of the yeast pet 56=his3-dedl gene region", Nucleic Acid Research, vol. 13, No. 23, 1985, pp.8587-8601.
Wolf et al, "Carboxypeptidase S-and Carboxypeptidase y-deficient mutants of Saccharomyces cerevisiae", p. 419, col. 1, pp. 424-425, vol. 147, No. 2, Aug. 1981, pp. 418-426.

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