Fig. 2

Detection of pMD18-O/MHRgfp in the transformed cells. M. hyorhinis were transformed with 15 µg pMD18-O/MHRgfp and grown in KM2 medium containing 0.01 µg/ml tetracycline hydrochloride. The same transformants were also plated in KM2-Agar containing 0.01 µg/ml tetracycline hydrochloride and the plate was incubated for 7 days. a Colonies of M. hyorhinis observed under microscopy (scale bar 100 µM). b Transformed mycoplasmas either subcultured in KM2 medium containing 0.01 µg/ml tetracycline hydrochloride or plated in KM2-Agar containing 0.01 µg/ml tetracycline hydrochloride and a single clone picked and sub-cultured in KM2 medium containing 0.01 µg/ml tetracycline hydrochloride. DNA was extracted from control un-transformed mycoplasmas or from transformed mycoplasmas and presence of pMD18-O/MHRgfp was evaluated by PCR to detect GFP from KM2 (b.A) or from KM2-Agar (b.B). The GFP product of about 750 bp was only detected from the DNA of the transformants mycoplasmas and it was consistent with the predicted sizes of GFP. This band was absent from the controls (un-transformed mycoplasmas)