Transposon Mutagenesis in Mycobacterium avium Subspecies Paratuberculosis

John P. Bannantine, Denise K. Zinniel, Raúl G. Barletta

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

While transposon mutagenesis has been developed for Mycobacterium avium subspecies paratuberculosis (Map), relatively few laboratories have adopted this important genetic tool to examine gene function and essentiality. Here we describe the construction of a Map transposon library using the Himar1 mariner transposon, but concepts can also be applied to the Tn5367 transposon, which has also been used by our group. Delivery of the transposon is by a temperature-sensitive phagemid, ϕMycoMarT7, and plating transductants requires patience and specialized media due to length of incubation required to observe colonies. Several transposon mutants obtained from these libraries have been tested in vaccine and pathogenesis studies. By providing the following detailed protocol herein, we expect to demystify the procedure and encourage additional investigators to incorporate transposon mutagenesis in their studies on Johne’s disease.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages117-125
Number of pages9
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume2016
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Johne’s disease
  • Mycobacterium
  • Transposon mutagenesis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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