Targeted chromosomal knockouts in mycoplasma pneumoniae

Radha Krishnakumar, Nacyra Assad-Garcia, Gwynedd A. Benders, Quang Phan, Michael G. Montague, John I. Glass

Research output: Contribution to journalArticle

Abstract

Most gene knockouts in mycoplasmas are achieved through labor-intensive transposon mutagenesis. Here, we describe a method for making targeted deletions in Mycoplasma pneumoniae by use of homologous recombination. In this method, M. pneumoniae is transformed with a plasmid carrying an antibiotic resistance marker flanked by 1-kb regions surrounding the target gene. Following selection for the antibiotic resistance, colonies are screened for double crossovers which indicate complete deletion of the target open reading frame.

Original languageEnglish (US)
Pages (from-to)5297-5299
Number of pages3
JournalApplied and environmental microbiology
Volume76
Issue number15
DOIs
StatePublished - Aug 1 2010

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

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    Krishnakumar, R., Assad-Garcia, N., Benders, G. A., Phan, Q., Montague, M. G., & Glass, J. I. (2010). Targeted chromosomal knockouts in mycoplasma pneumoniae. Applied and environmental microbiology, 76(15), 5297-5299. https://doi.org/10.1128/AEM.00024-10