Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes
Article Title: Activation of σ20-dependent recombination and horizontal gene transfer in Mycoplasma genitalium
Figure Lengend Snippet: Schematic representation of representative mating experiments. Qualitative assessment of recovery of double resistant mutants in mating experiments using M. genitalium strains carrying different antibiotic gene markers. (A) Mating experiments using donor strains (depicted as white mycoplasma cells) carrying the chloramphenicol (Cm R ) and tetracycline (Tet R ) resistance markers and recipient strains (depicted as light gray mycoplasma cells) carrying the puromycin (Pm R ) resistance marker. Donor strains overexpressing σ 20 are indicated as ‘Upσ 20 .’ One of the donor strains used, carried a deletion of the recA gene ( recA - ). Unsuccessful DNA transfers are indicated as empty, dash-lined mycoplasma cells. (B) Mating experiments using a previous transconjugant mutant carrying the chloramphenicol and puromycin resistance markers as a donor strain (depicted as a light gray mycoplasma cell) and a recipient strain (depicted as a dark gray mycoplasma cell) carrying the tetracycline resistance marker. Selection with tetracycline and chloramphenicol results in the isolation of a new transconjugant strain bearing the corresponding antibiotic markers and overexpressing σ 20 . Selection with puromycin and tetracycline allows the isolation of another transconjugant strain with the corresponding antibiotic markers but without the ectopic copy of σ 20 . All the transconjugant strains were genotyped by PCR and sequencing.
Article Snippet: Plasmids for M. genitalium transformation were obtained using the GenElute HP Midiprep kit (Sigma).
Techniques: Marker, Mutagenesis, Selection, Isolation, Polymerase Chain Reaction, Sequencing