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 journalArticlepeer-review

22 Scopus citations


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
Issue number15
StatePublished - Aug 2010
Externally publishedYes

ASJC Scopus subject areas

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


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