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. 2019 May 30;85(12):e00275-19. doi: 10.1128/AEM.00275-19

FIG 3.

FIG 3

Curing of the plasmid pLVM1 from Methylobacterium sp. strain DM1 and phenotype analysis. (A) Comparison of the growth abilities of DM1ΔpLVM1 and the wild-type strain on MSM agar plates containing 2,000 mg liter−1 DMF. Quadrants 1 and 4 represent the wild-type strain, and quadrants 2 and 3 represent the plasmid-free strain DM1ΔpLVM1. (B) Effects of eliminating plasmid pLVM1 from strain DM1 on growth and degradation of 2,000 mg liter−1 DMF in MSM. ▲, growth curve for DM1ΔpLVM1; ♦, DMF degradation curve for DM1ΔpLVM1; ■, growth curve for the wild-type strain; ●, DMF degradation curve for the wild-type strain. (C) PCR verification of plasmid curing. Genes dmfA1A2 and dmmD are plasmid-borne genes, and genes chr1 and chr2 are chromosomal genes. ΔP, DM1ΔpLVM1; WT, wild-type strain (positive control); M, molecular size markers. (D) Phenotype analysis of carbon or carbon and nitrogen source utilization of DM1ΔpLVM1 (−P), compared with the wild-type (WT) strain. The concentrations of substrates were as follows: 5 mg liter−1 DMA, 5 mg liter−1 MA, 3% (vol/vol) methanol, 3% (vol/vol) ethanol, 1% (vol/vol) toluene, 1% (vol/vol) formamide, 1% (vol/vol) glycerol, and 4% (vol/vol) trisodium citrate.