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Microbiology 152 (2006), 519-527; DOI  10.1099/mic.0.28287-0
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Microbiology 152 (2006), 519-527; DOI  10.1099/mic.0.28287-0
© 2006 Society for General Microbiology

Comparative analysis of antibiotic resistance gene markers in Mycoplasma genitalium: application to studies of the minimal gene complement

Oscar Q. Pich{dagger}, Raul Burgos{dagger}, Raquel Planell, Enrique Querol and Jaume Piñol

Institut de Biotecnologia i Biomedicina and Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain

Correspondence
Jaume Piñol
jaume.pinyol{at}uab.es

Mycoplasma genitalium has been proposed as a suitable model for an in-depth understanding of the biology of a free-living organism. This paper reports that the expression of the aminoglycoside resistance gene aac(6')-aph(2''), the only selectable marker hitherto available for M. genitalium genetic studies, correlates with a growth impairment of the resistant strains. In light of this finding, a tetM438 construction based on the tetracycline resistance gene tetM was developed; it can be used efficiently in M. genitalium and confers multiple advantages when compared to aac(6')-aph(2''). The use of tetM438 significantly improves transformation efficiency and generates visible colonies faster. Finally, the improvements in the pMTnTetM438 construction made it possible to obtain insertions in genes which have not been previously considered to be dispensable under laboratory growth conditions.

Abbreviations: CDA, colony diameter average; Gm, gentamicin; Tc, tetracycline

{dagger}These authors contributed equally to this work.




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