Targeted chromosomal knockouts in mycoplasma pneumoniae

Radha Krishnakumar, Nacyra Assad-Garcia, Gwynedd A. Benders, Quang Phan, Michael 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
Externally publishedYes

Fingerprint

Mycoplasma pneumoniae
pneumonia
antibiotic resistance
Microbial Drug Resistance
Gene Knockout Techniques
gene
Mycoplasma
Homologous Recombination
homologous recombination
gene targeting
plasmid
mutagenesis
Mutagenesis
transposons
Open Reading Frames
recombination
open reading frames
plasmids
labor
Plasmids

ASJC Scopus subject areas

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

Cite this

Krishnakumar, R., Assad-Garcia, N., Benders, G. A., Phan, Q., Montague, M., & 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

Targeted chromosomal knockouts in mycoplasma pneumoniae. / Krishnakumar, Radha; Assad-Garcia, Nacyra; Benders, Gwynedd A.; Phan, Quang; Montague, Michael; Glass, John I.

In: Applied and Environmental Microbiology, Vol. 76, No. 15, 01.08.2010, p. 5297-5299.

Research output: Contribution to journalArticle

Krishnakumar, R, Assad-Garcia, N, Benders, GA, Phan, Q, Montague, M & Glass, JI 2010, 'Targeted chromosomal knockouts in mycoplasma pneumoniae', Applied and Environmental Microbiology, vol. 76, no. 15, pp. 5297-5299. https://doi.org/10.1128/AEM.00024-10
Krishnakumar R, Assad-Garcia N, Benders GA, Phan Q, Montague M, Glass JI. Targeted chromosomal knockouts in mycoplasma pneumoniae. Applied and Environmental Microbiology. 2010 Aug 1;76(15):5297-5299. https://doi.org/10.1128/AEM.00024-10
Krishnakumar, Radha ; Assad-Garcia, Nacyra ; Benders, Gwynedd A. ; Phan, Quang ; Montague, Michael ; Glass, John I. / Targeted chromosomal knockouts in mycoplasma pneumoniae. In: Applied and Environmental Microbiology. 2010 ; Vol. 76, No. 15. pp. 5297-5299.
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