MX Cassettes for Knocking Out Genes in Yeast.

Published online

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • McCusker, JH

Published Date

  • April 3, 2017

Published In

Volume / Issue

  • 2017 / 4

Start / End Page

  • pdb.top080689 -

PubMed ID

  • 28373515

Pubmed Central ID

  • 28373515

Electronic International Standard Serial Number (EISSN)

  • 1559-6095

Digital Object Identifier (DOI)

  • 10.1101/pdb.top080689

Language

  • eng

Conference Location

  • United States