Table 1

Transplantation of M. mycoides YCp genomes from yeast into wild-type and RE(–) M. capricolum recipient cells. The number of tetracycline-resistant, blue colonies obtained after the transplantation of M. mycoides YCp genomes from yeast into M. capricolum recipients was counted. Wild-type M. capricolum and M. capricolum RE(–) transplantation was performed using methods described in fig. S1. For untreated samples, yeast plugs were digested with β-agarase (melting step) and transplanted into both recipient cells. The treated samples were methylated and treated with proteinase K before the melting step. The mock-methylated sample was treated the same as the methylated samples, except that no extract or purified methyltransferases were added. VL6-48N yeast agarose plugs used in this experiment carried YCpMmyc1.1. W303a yeast agarose plugs carried YCpMmyc1.1, YCpMmyc1.1 that was engineered in yeast (YCpMmyc1.1-ΔtypeIIIres::URA3 or YCpMmyc1.1-ΔtypeIIIres), or YCpMmyc1.1-Δ500kb. The number of transplants is the average of at least three experiments. The error reported is the absolute mean deviation.

Yeast strainGenomeMethylation treatmentNumber of transplants (colonies or plugs)
M. capricolum RE(–)Wild-type M. capricolum
VL6-48NYCpMmyc1.1Untreated37 ± 30
M. capricolum extracts32 ± 139 ± 4
M. mycoides extracts15 ± 822 ± 8 [13 ± 4]* [10 ± 4]†
Mock-methylated34 ± 170
M. mycoides purified methylases20 ± 1713 ± 10
W303aYCpMmyc1.1Untreated22 ± 5Not done
YCpMmyc1.1-ΔtypeIIIres::URA3Untreated52 ± 10Not done
YCpMmyc1.1-ΔtypeIIIresUntreated52 ± 12Not done
YCpMmyc1.1-Δ500kbUntreated0Not done

*Yeast plugs were cleared of yeast genomic DNA by digestion with a cocktail of Asi SI, Rsr II, and Fse I followed by pulsed-field gel electrophoresis.

†Yeast plugs were cleared of yeast genomic DNA by using pulsed-field gel electrophoresis.