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MX Cassettes for Knocking Out Genes in Yeast.

Publication ,  Journal Article
McCusker, JH
Published in: Cold Spring Harb Protoc
April 3, 2017

Precise modifications of the Saccharomyces cerevisiae genome use marker cassettes, most often in the form of "knockout" (KO) marker cassettes, to delete genes. Many different KO marker cassettes exist, some of which require strains with specific genotypes, such as auxotrophic mutations, and others that have no strain genotype requirements, such as selections for drug resistance and one of two selections for nitrogen source utilization. This introduction focuses on the most frequently used family of KO cassettes-the MX cassettes. In particular, we focus on and describe the different types of MX cassettes and selections; specifically, selections for prototrophy; selections for utilization of cytosine or acetamide as sole nitrogen sources; and selections for resistance to six different drugs. The use of cassettes to place genes under regulated control is briefly discussed. Also discussed are strain genotype requirements (where applicable); media requirements; how to "recycle" or "pop out" cassettes; and counterselections against specific KO cassettes.

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Published In

Cold Spring Harb Protoc

DOI

EISSN

1559-6095

Publication Date

April 3, 2017

Volume

2017

Issue

4

Start / End Page

pdb.top080689

Location

United States

Related Subject Headings

  • Selection, Genetic
  • Saccharomyces cerevisiae
  • Recombination, Genetic
  • Mutagenesis, Insertional
  • Microbiological Techniques
  • Gene Knockout Techniques
  • 1103 Clinical Sciences
  • 0601 Biochemistry and Cell Biology
  • 0306 Physical Chemistry (incl. Structural)
 

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McCusker, J. H. (2017). MX Cassettes for Knocking Out Genes in Yeast. Cold Spring Harb Protoc, 2017(4), pdb.top080689. https://doi.org/10.1101/pdb.top080689
McCusker, John H. “MX Cassettes for Knocking Out Genes in Yeast.Cold Spring Harb Protoc 2017, no. 4 (April 3, 2017): pdb.top080689. https://doi.org/10.1101/pdb.top080689.
McCusker JH. MX Cassettes for Knocking Out Genes in Yeast. Cold Spring Harb Protoc. 2017 Apr 3;2017(4):pdb.top080689.
McCusker, John H. “MX Cassettes for Knocking Out Genes in Yeast.Cold Spring Harb Protoc, vol. 2017, no. 4, Apr. 2017, p. pdb.top080689. Pubmed, doi:10.1101/pdb.top080689.
McCusker JH. MX Cassettes for Knocking Out Genes in Yeast. Cold Spring Harb Protoc. 2017 Apr 3;2017(4):pdb.top080689.

Published In

Cold Spring Harb Protoc

DOI

EISSN

1559-6095

Publication Date

April 3, 2017

Volume

2017

Issue

4

Start / End Page

pdb.top080689

Location

United States

Related Subject Headings

  • Selection, Genetic
  • Saccharomyces cerevisiae
  • Recombination, Genetic
  • Mutagenesis, Insertional
  • Microbiological Techniques
  • Gene Knockout Techniques
  • 1103 Clinical Sciences
  • 0601 Biochemistry and Cell Biology
  • 0306 Physical Chemistry (incl. Structural)