T. Hahn et al., Construction and analysis of a modified Tn4001 conferring chloramphenicol resistance in Mycoplasma pneumoniae, PLASMID, 41(2), 1999, pp. 120-124
The Staphylococcus aureus transposon Tn4001 and derivatives thereof have be
en transformed successfully in several mycoplasma species. In order to expa
nd the versatility of Tn4001 for other genetic manipulations and for use in
mycoplasma species resistant to gentamicin (Gm), chloramphenicol acetyltra
nsferase (Cat) from S. aureus was evaluated as a selectable marker. The cat
gene was cloned in both orientations into a modified Tn4001 and transforme
d into Mycoplasma pneumoniae, conferring resistance to Cm and Gm. Replaceme
nt of the gene for Gm(R) in Tn4001 with cut likewise conferred Cm-R when tr
ansformed into M. pneumoniae. The minimum inhibitory concentration to Cm in
transformants with cat derivatives of Tn4001 was 300-500 mu g/ml, and Cat
enzyme activity was demonstrated by using a fluorescent substrate. (C) 1999
Academic Press.