Figure 4.

Western blot analysis of Val1686 secretion. Overnight cultures of Δval1686 strain and several complemented strains. Complementation included strains that synthesis native Val1686 (pval1686) or Val1686 without the first 30 amino acids (pval1686Δ30; negative control), or Val1686 having an amino acid change (histidine to alanine, position 348; pval1686 H348A) or Val1686 lacking the Fic domain (pval1686ΔFic). Equal volume (10 μL) of prepared samples were loaded onto SDS-PAGE gel and the histidine tag in C-terminus was probed for individual recombinant protein in western blot. Anti-DnaK was used as a loading control. The data are representative of three repeated experiments.