Journal: BMC Microbiology

Article Title: The conserved actinobacterial transcriptional regulator FtsR controls expression of ftsZ and further target genes and influences growth and cell division in Corynebacterium glutamicum

doi: 10.1186/s12866-019-1553-0

Figure Lengend Snippet: Growth behavior and cell morphology of C. glutamicum Δ ftsR and C. glutamicum overexpressing ftsR . a Growth of the ftsR deletion mutant in comparison to the wild type. Mean values and standard deviation of three biological replicates are shown. b Growth of an ftsR overexpressing strain and complementation of the Δ ftsR mutant with several FtsR variants including an N-terminally shortened protein and a protein with a C-terminal StrepTag-II. Average values from two biological replicates are shown. For the growth experiments shown in a and b , the strains were pre-cultivated first in BHI medium and then in CGXII medium with 2% (w/v) glucose, followed by the main cultivation in the same medium. For b , all media were supplemented with kanamycin (25 μg/mL) and IPTG as indicated. c Microscopic pictures of cells in the stationary phase. To visualize membranes and DNA, cells were stained with Nile Red and Hoechst 33342, respectively. The arrow points towards a branched cell and the circle indicates a high DNA concentration at the cell pole. The scale bar represents 5 μm

Article Snippet: For growth experiments, C. glutamicum was precultivated for 6–8 h at 30 °C and 170 rpm in 5 mL BHI medium (BD BactoTM Brain Heart Infusion, Becton Dickinson and Company, Heidelberg, Germany).

Techniques: Mutagenesis, Comparison, Standard Deviation, Staining, Concentration Assay

Journal: BMC Microbiology

Article Title: The conserved actinobacterial transcriptional regulator FtsR controls expression of ftsZ and further target genes and influences growth and cell division in Corynebacterium glutamicum

doi: 10.1186/s12866-019-1553-0

Figure Lengend Snippet: Promoter exchange of ftsZ and growth of the resulting strains. a Strains with FtsR-independent ftsZ expression were constructed using a DNA fragment with a terminator sequence and the gluconate-inducible gntK promoter, which was inserted between the native ftsZ promoter and the ftsZ start codon in the chromosomes of MB001 and the MB001Δ ftsR mutant. b-d Effect of different gluconate concentrations on cell morphology ( b ) and growth ( c, d ) of the promoter exchange strains MB001::P gntK - ftsZ and MB001Δ ftsR ::P gntK - ftsZ . The two strains were first pre-cultivated in BHI medium supplemented with 0.1% (w/v) gluconate to induce ftsZ expression by P gntK . The second pre-cultivation was performed in CGXII medium with 2% (w/v) glucose supplemented with the indicated gluconate concentrations. The main cultures were then performed in media having the same composition as the ones for the second pre-cultivation. b Microscopic pictures of cells from the stationary phase. The scale bar represents 5 μm. c, d The growth experiments show mean values and standard deviations of three biological replicates

Article Snippet: For growth experiments, C. glutamicum was precultivated for 6–8 h at 30 °C and 170 rpm in 5 mL BHI medium (BD BactoTM Brain Heart Infusion, Becton Dickinson and Company, Heidelberg, Germany).

Techniques: Expressing, Construct, Sequencing, Mutagenesis