FIG. 1.

Physical map of the dnaK region in wild-type strain MG1363 and dnaK mutants and Western blot analysis of mutant proteins. (A and B) Gray boxes represent structural genes, and the gene designation is indicated. (A) Physical map of the hrcA-grpE-dnaK operon in MG1363. (B) Extent of dnaK sequences in the hrcA-grpE-dnaK operon in mutant strains BK6 and BK11, shown by lines below a partial restriction map of the dnaK gene. The mutants were constructed by homologous recombination into the chromosome of various plasmids (see text for details). (C) Functional domains recognized for the DnaK protein from E. coli and the eucaryotic DnaK homologs Hsp70. (D) Western blot analysis of mutant proteins with antibodies against DnaK from B. subtilis. Proteins were extracted from exponentially growing cells and separated by SDS-PAGE. Following transfer of the protein bands to an Immobilon-P membrane, the DnaK protein was detected by chemiluminescence with an Amersham ECL Western blotting analysis system and antibodies against DnaK from B. subtilis. The approximate sizes of the proteins are shown above the bands.