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

1996-11-15

1999-12-21

Degen, Nancy

Chemistry: molecular biology and microbiology

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

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

4351721, 43525421, 4353201, C12P 2102, C12N 115, C12N 116, C12N 1581

Patent

active

060047763

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, Rhizopus and Trichoderma, the expression vector containing both an "expressible gene" encoding the protein and a so-called "deficient selection marker needed for the growth of the yeast or mould in a specific medium", such as the LEU2d, TRP1d or URA3d gene, in combination with a 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, "Heterologous Gene Expression in Saccharmyces cerevisiae", Biotech. Gen. Eng. Rev. 12, 377-406.
Lopes et al, "High-Copy-Number Integration In the Ribosomal . . . ", Helsinki, 1988 (abstract).
Lopes et al, "High-copy-number integration into the ribosomal . . . ", Gene, 79 (1989) 199-206.

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