TABLE 6.

MinD stimulates division inhibition by the membrane-tethered ZipA′-GFP-MinC fusion protein

PhageaFusion proteinPlasmida (genotype)Relative level of fusion proteinbLoccLength (μm)ne
AvgRanged
λPC100ZipA′-GFP-MinCpDB3261.0M18.31.1-87.0135
pCH204 (minD)0.9M44.72.7-132.1134
λLL49ZipA′-GFP-DMinCpDB3261.1M4.21.5-11.5116
pCH204 (minD)1.0R6.01.7-25.8129
  • a Cells of strain DR109 (ΔminCDE recA), harboring the indicated phage and plasmid, were grown at 30°C to an OD600 of 0.12 to 0.14 in minimal medium supplemented with 50 μM IPTG. pDB326 was the parent vector.

  • b Protein levels were measured by quantitative Western analyses, using anti-GFP antibodies. Values are normalized to that of the ZipA′-GFP-MinC fusion in DR109(λPC100)/pDB326. The level of ZipA′-GFP-MinC in these cells corresponded to approximately four times the level of native MinC in strain CH3 (recA) as determined by quantitative Western analyses, using anti-MinC antibodies.

  • c The localization (Loc) of fusion proteins, and the average cell lengths, were determined by microscopy. M, concentrated along the membrane; R, accumulated in rings.

  • d Note that cultures of DR109 normally contain a small fraction of filaments due to the ΔminCDE and recA mutations in this strain.

  • e Number of cells measured.