Strains of pichia pastoris created by interlocus recombination

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

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435254, 435255, 435256, 935 28, 935 97, C12N 1500, C12N 114, C12N 116

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051224658

ABSTRACT:
A process for the regeneration of a selectable phenotype at a high frequency by interlocus recombination, in yeast cultures transformed with recombinant DNA constructs comprised of a gene whose product complements the selectable phenotype to be regenerated is disclosed. A sequence of steps is involved including; suspending together in rich media mutant yeast strains, plating onto presporulation agar, replica plating onto other agar media suitable for selection and growth of diploid cells, plating on sporulation agar or alternatively dissecting the asci, digesting the cells, growing the spores on minimal media, and screening for the desired phenotype.

REFERENCES:
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H. Klein & T. Petes: Intrachromosomal Gene Conversion in Yeast; Nature; 289, 144-148 (1981).
J. Jackson & G. Fink: Meiotic Recombination Between Duplicated Genetic Elements in Saccharomyces Cerevisiae; Genetics 109, pp. 303-332 (1985).
S. Jinks-Robinson & T. Petes, High-Frequency Meiotic Gene Conversion Between Repeated Genes on Nonhomologous Chromosomes in Yeast; Proc. Natl. Acad. Sci., USA, 82, pp. 3350-3354 (1985).
Hybridization and Genetic Analysis of the Methanol Yeasts Pichia pinus; Genetika, 13, No. 2, pp. 322-329, (1976).
J. Jackson & G. Fink, Gene Conversion Between Duplicated Genetic Elements in Yeast; Nature 292, 306-311 (1981).
H. Rudolph et al., One-Step Gene Replacement in Yeast by Cotransformation; Gene 36, 87-85, (1985).
A Method for Gene Disruption . . . Alani et al., Genetics 116:541-545 (1987).
Functional Characterization of Two Alcohol Oxidase Genes, Cregg et al., Molecular & Cellular Bio., vol. 9, No. 3, pp. 1316-1323 (1989).

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