TY - JOUR
T1 - Targeted chromosomal knockouts in mycoplasma pneumoniae
AU - Krishnakumar, Radha
AU - Assad-Garcia, Nacyra
AU - Benders, Gwynedd A.
AU - Phan, Quang
AU - Montague, Michael G.
AU - Glass, John I.
N1 - Copyright:
Copyright 2010 Elsevier B.V., All rights reserved.
PY - 2010/8
Y1 - 2010/8
N2 - 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.
AB - 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.
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U2 - 10.1128/AEM.00024-10
DO - 10.1128/AEM.00024-10
M3 - Article
C2 - 20543037
AN - SCOPUS:77955574465
SN - 0099-2240
VL - 76
SP - 5297
EP - 5299
JO - Applied and environmental microbiology
JF - Applied and environmental microbiology
IS - 15
ER -