Preparation of Pichia methanolica auxotrophic mutants

Details Preparation of Pichia methanolica auxotrophic mutants Preparation of Pichia methanolica auxotrophic mutants

1996-08-26

1998-04-07

Saunders, David

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Fungi

435 34, 4351721, 4352555, 935 37, C12N 116, C12N 1501

Patent

active

057363838

ABSTRACT:
Methods for preparing Pichia methanolica cells having auxotrophic mutations are disclosed. The methods comprise the steps of (a) exposing P. methanolica cells to mutagenizing conditions, (b) culturing the cells from step (a) in a rich medium to allow mutations to become established and replicated in at least a portion of the cells, (c) culturing the cells from step (b) in a culture medium deficient in assimilable nitrogen to deplete cellular nitrogen stores, (d) culturing the cells from step (c) in a defined culture medium comprising an inorganic nitrogen source and an amount of nystatin sufficient to kill growing P. methanolica cells to select for cells having a deficiency in a nutritional gene; and (e) culturing the selected cells from step (d) in a rich culture medium.

REFERENCES:
patent: 4879231 (1989-11-01), Stroman et al.
American Type Culture Collection. Catalogue of Yeasts: Eighteenth edition, 1990. Jong, SC and Edwards, MJ, eds. Rockville, MD, 1990.
Janatova, I. Antoine van Leeuwenhoek. 62: 167-171, 1992.
Gould et al., Yeast 8: 613-628, 1992.
Piredda et al., Yeast 10: 1601-1612, 1994.
Hiep et al., Yeast 9: 1251-1258, 1993.
Strathem (ed.) et al., The Molecular Biology of the Yeast Saccharomyces: 271-279, 1982.
Hiep et al., Yeast 9: 1189-1197, 1993.

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