Fig 4.

Growth of C. glutamicum WT (open circles), C. glutamicum IMcg2701 (open triangles), C. glutamicum IMcg2701(pXMJ19-cg2701) (filled triangles), C. glutamicum IMcg2701(pXMJ19-cg2701-strep) (open squares), and C. glutamicum IMcg2701(pXMJ19) (filled circles) in CgC minimal medium with 2% (wt/vol) maltose (A), analysis of the cellular localization of MusI-Strep (B), and topology model of MusI (C). For localization of MusI-Strep, C. glutamicum IMcg2701(pXMJ19-cg2701-strep) cells were harvested, washed, and disrupted. Cytosolic and membrane fractions separated by ultracentrifugation were then electrophoresed on a 12% SDS-polyacrylamide gel and transferred to a membrane. MusI-Strep was detected using anti-Strep antibody. A PAGE ruler prestained protein ladder (MBI Fermentas) was used as the marker. The topology model of the membrane-spanning regions of MusI is based on topology modeling. Membrane-spanning sequences were determined based on consensus sequences from two topology prediction algorithms. One representative growth curve from at least three independent cultivations is shown. The results of each of the cultivations were comparable.