Process for preparing a protein by a fungus transformed by multi

Details Process for preparing a protein by a fungus transformed by multi Process for preparing a protein by a fungus transformed by multi

1994-12-27

2000-07-18

Fleisher, Mindy

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435 691, 4351721, C12P 2106

Patent

active

060905743

ABSTRACT:
A process is disclosed for preparing a protein by a eukaryote transformed by multicopy integration of an expression vector into the genome of a yeast, such as Saccharomyces, Hansenula, and Kluyveromyces, or of a mould such as Aspergillus, Rbizopus and Trichoderma, the expression vector containing both an "expressible gene" encoding the protein and a so-called "deficient selection marker meeded for the growth of the yeast or mould in a specific medium", such as the LEU2d, TRP1d or URA3d gene, in combination witha ribosomal DNA sequence, resulting in stable high copy integration of 100-300 copies per cell. This multicopy integration results in an increased production of the desired protein, which can be guar .alpha.-galactosidase, an oxidase or a hydrolytic enzyme such as a lipase.

REFERENCES:
patent: 4935496 (1990-06-01), Kudo et al.
patent: 5001230 (1991-03-01), Brown et al.
Kingsman et al, "Heterologus Gene Expression in Saccharmyces cerevisiae", Biotech.Gen.Eng.Rev. 12, 377-406 1985.
Lopes et al., "High-Copy-Number Intergration in The Ribosomal . . . ", Helsinki, 1988 (Abstract).
Lopes et al., "High-copy-number intergration into the ribosomal . . . ", Gene, 79 (1989) 199-206.

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