Gene essentiality testing in mycobacterium smegmatis using specialized transduction

Apoorva Bhatt, William R. Jacobs

Research output: Chapter in Book/Report/Conference proceedingChapter

6 Scopus citations

Abstract

Conditional expression-specialized transduction essentiality test (CESTET) is a genetic tool used to determine essentiality of individual genes in Mycobacterium smegmatis. CESTET combines specialized transduction, a highly efficient gene knockout method, with the utility of the inducible acetamidase promoter. In a merodiploid strain containing a second integrated copy of an essential gene under the control of the acetamidase promoter, transductants (gene knockouts of the native chromosomal copy of a gene) are obtained only in the presence of acetamide in the selection medium. Furthermore, effects of loss of essential gene function can then be studied by growing the transductants in medium depleted of acetamide.

Original languageEnglish (US)
Title of host publicationMycobacteria Protocols
Subtitle of host publicationSecond Edition
PublisherHumana Press Inc.
Pages325-336
Number of pages12
ISBN (Print)9781588298898
DOIs
StatePublished - Jan 1 2008

Publication series

NameMethods in Molecular Biology
Volume465
ISSN (Print)1064-3745

Keywords

  • Acetamidase
  • Conditional expression
  • Essentiality testing
  • Phages
  • Specialized transduction

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Bhatt, A., & Jacobs, W. R. (2008). Gene essentiality testing in mycobacterium smegmatis using specialized transduction. In Mycobacteria Protocols: Second Edition (pp. 325-336). (Methods in Molecular Biology; Vol. 465). Humana Press Inc.. https://doi.org/10.1007/978-1-59745-207-6_22