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Reference - PMID:17622533 - Six new amino acid-auxotrophic markers for targeted gene integration and disruption in fission yeast.

Reference summary

PubMed ID
PMID:17622533
Title
Six new amino acid-auxotrophic markers for targeted gene integration and disruption in fission yeast.
Authors
Ma Y, Sugiura R, Saito M, Koike A, Sio SO, Fujita Y, Takegawa K, Kuno T
Citation
Curr Genet 2007 Aug;52(2):97-105
Publication year
2007
Abstract
Fission yeast Schizosaccharomyces pombe is amenable to genetics and is an excellent model system for studying eukaryotic cell biology. However, auxotrophic markers that can be used for both targeted gene integration and disruption are very limited. Here we performed a forward genetic screen in an effort to develop a new set of selectable markers for use in this yeast. Mutants that were auxotrophic for arginine, asparagine, cysteine, lysine, methionine and phenylalanine were isolated. Six genes were analyzed in detail and the mutations in the genes were identified. Among these six are three new genes: asn1 (+), cys2 (+) and pha2 (+) were required for biosynthesis of asparagine, cysteine and phenylalanine, respectively. New alleles of arg1 (+), lys3 (+) and met6 (+) were also identified. All of these genes proved to be suitable as selectable markers for targeted gene integration and disruption. We also showed that in Schizosaccharomyces pombe there are two apparent homologues of Saccharomyces cerevisiae MET2: the previously known met6 (+), and SPBC106.17c (named cys2 (+)). The cys2 mutation required cysteine rather than methionine. These new tools, specifically, new selectable markers, will be useful in further genetic and biological studies in fission yeast.

Annotation

Experimental tools

PBO:0003176 - selectable marker, amino acid auxotroph

Genes:

GO biological process

GO:0006526 - L-arginine biosynthetic process

Genes:

GO:0070981 - L-asparagine biosynthetic process

Genes:

GO:0019344 - L-cysteine biosynthetic process

Genes:

GO:0019878 - L-lysine biosynthetic process via aminoadipic acid

Genes:

GO:0071265 - L-methionine biosynthetic process

Genes:

GO:0009094 - L-phenylalanine biosynthetic process

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Single locus phenotype

FYPO:0001407 - decreased cell population growth on glucose carbon source

Genes:

Genotypes:

FYPO:0000035 - growth auxotrophic for arginine

Genes:

Genotypes:

FYPO:0000036 - growth auxotrophic for asparagine

Genes:

Genotypes:

FYPO:0000037 - growth auxotrophic for cysteine

Genes:

Genotypes:

FYPO:0000039 - growth auxotrophic for lysine

Genes:

Genotypes:

FYPO:0000040 - growth auxotrophic for methionine

Genes:

Genotypes:

FYPO:0000041 - growth auxotrophic for phenylalanine

Genes:

Genotypes: