Designer deletion and prototrophic strains derived from Saccharomyces cerevisiae strain W303-1a

Kirstin Replogle; Laura Hovland; David H. Rivier

doi:10.1002/(SICI)1097-0061(199908)15:11<1141::AID-YEA439>3.0.CO;2-P

Designer deletion and prototrophic strains derived from Saccharomyces cerevisiae strain W303-1a

Kirstin Replogle, Laura Hovland, David H. Rivier

Cell and Developmental Biology

Research output: Contribution to journal › Article › peer-review

Abstract

We report the construction of Saccharomyces cerevisiae strains isogenic to W303-1a that are designed to allow efficient genetic analysis. To facilitate the generation of null alleles of target genes by PCR-mediated gene disruption, we constructed designer deletion alleles of the ARG4, TRP1 and URA3 genes. In addition, a single pair of oligonucleotide primers were designed that can be used to amplify any of several marker genes for use in PCR-mediated gene disruption. A new version of the 'reusable' hisG-URA3-hisG cassette was constructed for use in PCR-mediated gene disruption. Finally, to facilitate the formation of isogenic diploids by selection, we constructed strains that contain combinations of wild-type alleles of ADE2, HIS3, LEU2, TRP1 and URA3.

Original language	English (US)
Pages (from-to)	1141-1149
Number of pages	9
Journal	Yeast
Volume	15
Issue number	11
DOIs	https://doi.org/10.1002/(SICI)1097-0061(199908)15:11<1141::AID-YEA439>3.0.CO;2-P
State	Published - 1999

Keywords

ARG4
Auxotrophic marker
Gene disruption
Saccharomyces cerevisiae
W303
Yeast

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biochemistry
Applied Microbiology and Biotechnology
Genetics

Online availability

10.1002/(SICI)1097-0061(199908)15:11<1141::AID-YEA439>3.0.CO;2-P

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abstract = "We report the construction of Saccharomyces cerevisiae strains isogenic to W303-1a that are designed to allow efficient genetic analysis. To facilitate the generation of null alleles of target genes by PCR-mediated gene disruption, we constructed designer deletion alleles of the ARG4, TRP1 and URA3 genes. In addition, a single pair of oligonucleotide primers were designed that can be used to amplify any of several marker genes for use in PCR-mediated gene disruption. A new version of the 'reusable' hisG-URA3-hisG cassette was constructed for use in PCR-mediated gene disruption. Finally, to facilitate the formation of isogenic diploids by selection, we constructed strains that contain combinations of wild-type alleles of ADE2, HIS3, LEU2, TRP1 and URA3.",

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AU - Replogle, Kirstin

AU - Hovland, Laura

AU - Rivier, David H.

PY - 1999

Y1 - 1999

N2 - We report the construction of Saccharomyces cerevisiae strains isogenic to W303-1a that are designed to allow efficient genetic analysis. To facilitate the generation of null alleles of target genes by PCR-mediated gene disruption, we constructed designer deletion alleles of the ARG4, TRP1 and URA3 genes. In addition, a single pair of oligonucleotide primers were designed that can be used to amplify any of several marker genes for use in PCR-mediated gene disruption. A new version of the 'reusable' hisG-URA3-hisG cassette was constructed for use in PCR-mediated gene disruption. Finally, to facilitate the formation of isogenic diploids by selection, we constructed strains that contain combinations of wild-type alleles of ADE2, HIS3, LEU2, TRP1 and URA3.

AB - We report the construction of Saccharomyces cerevisiae strains isogenic to W303-1a that are designed to allow efficient genetic analysis. To facilitate the generation of null alleles of target genes by PCR-mediated gene disruption, we constructed designer deletion alleles of the ARG4, TRP1 and URA3 genes. In addition, a single pair of oligonucleotide primers were designed that can be used to amplify any of several marker genes for use in PCR-mediated gene disruption. A new version of the 'reusable' hisG-URA3-hisG cassette was constructed for use in PCR-mediated gene disruption. Finally, to facilitate the formation of isogenic diploids by selection, we constructed strains that contain combinations of wild-type alleles of ADE2, HIS3, LEU2, TRP1 and URA3.

KW - ARG4

KW - Auxotrophic marker

KW - Gene disruption

KW - Saccharomyces cerevisiae

KW - W303

KW - Yeast

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IS - 11

ER -

Designer deletion and prototrophic strains derived from Saccharomyces cerevisiae strain W303-1a

Abstract

Keywords

ASJC Scopus subject areas

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