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Large‐scale transposon mutagenesis of Mycoplasma pulmonis
Author(s) -
French Christopher T.,
Lao Ping,
Loraine Ann E.,
Matthews Brian T.,
Yu Huilan,
Dybvig Kevin
Publication year - 2008
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2008.06262.x
Subject(s) - biology , transposable element , transposon mutagenesis , mutagenesis , genetics , mutant , gene , insertional mutagenesis , bacillus subtilis , mycoplasma , phenotype , dna , transposition (logic) , dna repair , microbiology and biotechnology , bacteria , linguistics , philosophy
Summary To obtain mutants for the study of the basic biology and pathogenic mechanisms of mycoplasmas, the insertion site of transposon Tn 4001T was determined for 1700 members of a library of Mycoplasma pulmonis mutants. After evaluating several criteria for gene disruption, we concluded that 321 of the 782 protein coding regions were inactivated. The dispensable and essential genes of M. pulmonis were compared with those reported for Mycoplasma genitalium and Bacillus subtilis . Perhaps the most surprising result of the current study was that unlike other bacteria, ribosomal proteins S18 and L28 were dispensable. Carbohydrate transport and the susceptibility of selected mutants to UV irradiation were examined to assess whether active transposition of Tn 4001T within the genome would confound phenotypic analysis. In contrast to earlier reports suggesting that mycoplasmas were limited in their DNA repair machinery, mutations in recA , uvrA , uvrB and uvrC resulted in a DNA‐repair deficient phenotype. A mutant with a defect in transport of N ‐acetylglucosamine was identified.