TasA production and pre-TasA processing during growth
and sporulation. (A) Samples of DSM cultures of strain AH94
(gerE36) were harvested during the logarithmic phase of
growth (lane 1), at T0 (lane 2),
T2 (lane 3), T4 (lane 4),
and T6 (lane 5), and whole-cell extracts were
prepared. The cultures were allowed to sporulate, and coat material was
then isolated from spores purified 24 h after
T0 (lane 6). DSM cultures of a ςH
mutant (strain AH17; lane 7) or of a gerE tasA insertional
mutant (strain AH1827, lane 8) were also collected at
T2. (B) Samples of supernatants of a culture of
a tasA+ strain (AH94, lane 1) or of a
tasA mutant (AH1700, lane 2) were collected at
T2, and the proteins were concentrated as
described in Materials and Methods. (C) The inducer IPTG was added to a
DSM culture of the Pspac-tasA strain AH1802 at
T0. Samples were collected at 30 (lane 1), 60
(lane 2), 90 (lane 3), and 120 min (lane 4) after the addition of IPTG,
and whole-cell lysates were prepared. Samples of DSM cultures of a
tasA mutant (lane 5) and of a tasA+
strain (lane 6) at T2 were also analyzed.
Proteins in all samples were resolved on 12.5% polyacrylamide gels
containing SDS and transferred to nitrocellulose membranes. The
membranes were probed with an anti-TasA antiserum. The arrows indicate
the positions of the TasA antigen. Molecular mass markers (in
kilodaltons) are also indicated.