The growth rate of Mycobacterium smegmatis depends on sufficient porin-mediated influx of nutrients
Version of Record online: 28 SEP 2005
Volume 58, Issue 3, pages 714–730, November 2005
How to Cite
Stephan, J., Bender, J., Wolschendorf, F., Hoffmann, C., Roth, E., Mailänder, C., Engelhardt, H. and Niederweis, M. (2005), The growth rate of Mycobacterium smegmatis depends on sufficient porin-mediated influx of nutrients. Molecular Microbiology, 58: 714–730. doi: 10.1111/j.1365-2958.2005.04878.x
- Issue online: 28 SEP 2005
- Version of Record online: 28 SEP 2005
- Accepted 22 August, 2005.
Table S1. Plasmids used in this work. Up- and downstream homologous sequences of genes are subscripted as up and down. ?Origin? means origin of replication. The annotations AmpR, HygR and KanR indicate that the plasmid confers resistance to ampicillin, hygromycin and kanamycin, respectively.
Table S2. Oligonucleotides used in this work. Restriction sites are underlined. The consensus ribosome binding site in the oligonucleotides for the msp expression vectors is marked in bold. Oligonucleotides mspBdownf and mspBdownr were used for the mspB probe and CS4-02 and CS4-09 for the mspD probe.
Table S3. Strains used in this work. The annotations SmR, GenR and HygR indicate that the strain is resistant to the antibiotics streptomycin, gentamicin and hygromycin, respectively. It should be noted that all strains are derivatives of SMR5 and are therefore resistant to streptomycin. This is not indicated in the table except for SMR5.
Fig. S1. Uptake of glucose and glycerol by the porin triple mutant M. smegmatis ML16. The accumulation of 20?μM [14C]glucose (A) and 20?μM [14C]glycerol (B) by the porin double mutant M.?smegmatis ML10 (ΔmspA ΔmspC; black circles) and the porin triple mutant M.?smegmatis ML16 (ΔmspA ΔmspC ΔmspD; white circles) was measured at 37°C for 16 minutes. The experiments were done in triplicate. Data are shown with standard deviations.
Fig. S2. Complementation of the growth defect of the porin double mutant M.?smegmatis ML10. M.?smegmatis SMR5/pMS2 (open circles), MN01/pMN016 (closed circles) and ML10/pMN016 (closed squares) were grown in 7H9 medium containing 0.2% glycerol (22?mM). The experiments were done in triplicate. Data are shown with standard deviations. Generation times were determined by regression analysis of the exponential phase (OD600 ? 1 to 4) for all strains.
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