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Heterologous URA3MX cassettes for gene replacement in Saccharomyces cerevisiae.

Publication ,  Journal Article
Goldstein, AL; Pan, X; McCusker, JH
Published in: Yeast
April 1999

Heterologous gene replacement cassettes are powerful tools for dissecting gene function in Saccharomyces cerevisiae. Their primary advantages over homologous gene replacement cassettes include reduced gene conversion (leading to efficient site-specific integration of the cassette) and greater independence of strain background. Perhaps the most widely used cassettes are the MX cassettes containing the dominant selectable kanamycin resistance gene (kanr), which confers resistance to G418 (Wach et al., 1994). One limitation of the kanMX cassettes is that they are not counterselectable and therefore not readily recyclable, which is important when constructing strains with more than one gene deletion. To address this limitation, and to expand the choices of heterologous markers, we have created two new MX cassettes by replacing the kanr ORF from plasmids pFA6-kanMX3 and pFA6-kanMX4 with the Candida albicans URA3 ORF. These plasmids, pAG60 (CaURA3MX4) and pAG61 (CaURA3MX3) are identical to the kanMX cassettes in all other respects but have the added advantage of being counterselectable and therefore readily recyclable in S. cerevisiae.

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Published In

Yeast

DOI

ISSN

0749-503X

Publication Date

April 1999

Volume

15

Issue

6

Start / End Page

507 / 511

Location

England

Related Subject Headings

  • Sequence Homology, Nucleic Acid
  • Selection, Genetic
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae
  • Recombination, Genetic
  • Plasmids
  • Orotic Acid
  • Open Reading Frames
  • Mutagenesis, Insertional
  • Kanamycin
 

Citation

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Goldstein, A. L., Pan, X., & McCusker, J. H. (1999). Heterologous URA3MX cassettes for gene replacement in Saccharomyces cerevisiae. Yeast, 15(6), 507–511. https://doi.org/10.1002/(SICI)1097-0061(199904)15:6<507::AID-YEA369>3.0.CO;2-P
Goldstein, A. L., X. Pan, and J. H. McCusker. “Heterologous URA3MX cassettes for gene replacement in Saccharomyces cerevisiae.Yeast 15, no. 6 (April 1999): 507–11. https://doi.org/10.1002/(SICI)1097-0061(199904)15:6<507::AID-YEA369>3.0.CO;2-P.
Goldstein AL, Pan X, McCusker JH. Heterologous URA3MX cassettes for gene replacement in Saccharomyces cerevisiae. Yeast. 1999 Apr;15(6):507–11.
Goldstein, A. L., et al. “Heterologous URA3MX cassettes for gene replacement in Saccharomyces cerevisiae.Yeast, vol. 15, no. 6, Apr. 1999, pp. 507–11. Pubmed, doi:10.1002/(SICI)1097-0061(199904)15:6<507::AID-YEA369>3.0.CO;2-P.
Goldstein AL, Pan X, McCusker JH. Heterologous URA3MX cassettes for gene replacement in Saccharomyces cerevisiae. Yeast. 1999 Apr;15(6):507–511.
Journal cover image

Published In

Yeast

DOI

ISSN

0749-503X

Publication Date

April 1999

Volume

15

Issue

6

Start / End Page

507 / 511

Location

England

Related Subject Headings

  • Sequence Homology, Nucleic Acid
  • Selection, Genetic
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae
  • Recombination, Genetic
  • Plasmids
  • Orotic Acid
  • Open Reading Frames
  • Mutagenesis, Insertional
  • Kanamycin