Localization of CwlJ within spores
Polyclonal antibodies were raised against overexpressed and purified histidinetagged
CwlJ, SleB and YpeB proteins, and were used in Western blotting
experiments. The SleB protein was overexpressed in a form that corresponded
to the processed, mature form of the enzyme; attempts to overexpress the
protein complete with its signal sequence failed. Cross-reacting anti-E. coli
antibodies were reduced or removed by purification on a column containing
immobilized E. coli lysate (Pierce). Each antibody preparation was able to
recognize its cognate protein antigen overexpressed in E. coli (data not
shown). Spores of strain HR, the laboratory wild-type 168 strain, or of
otherwise isogenic mutants, were used.