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. 1999 May;181(10):3010–3017. doi: 10.1128/jb.181.10.3010-3017.1999

FIG. 2.

FIG. 2

Overexpression of the SecYEG complex does not improve export in secBL75Q mutant strains. Cells were grown in minimal arabinose medium supplemented with ampicillin and a mix of 18 amino acids. (A) Immunoblots of total SecY and SecE protein. Samples (1 ml) were precipitated with trichloroacetic acid and analyzed by high-Tris SDS-PAGE (3). Proteins were transferred to a PVDF membrane and probed with anti-SecY and anti-SecE antisera. (B) Pulse-chase analysis of OmpA export. Cells were labeled with Tran35S-label for 15 s, and the label was chased with nonradioactive methionine and chloramphenicol. Samples were removed at the times indicated and treated as for Fig. 1, except that the samples were immunoprecipitated with anti-OmpA antiserum. Samples are as follows: lane a, HAC151 (secB+/pBAD22), 15-s pulse; lane b, HAC151, 0.5-min chase; lane c, HAC151, 1.0-min chase; lane d, HAC97 (secB+/pHAsecEYG), 15-s pulse; lane e, HAC97, 0.5-min chase; lane f, HAC97, 1.0-min chase; lane g, HAC152 (secBL75Q/pBAD22), 15-s pulse; lane h, HAC152, 0.5-min chase; lane i, HAC152, 1.0-min chase; lane j, HAC98 (secBL75Q/pHAsecEYG), 15-s pulse; lane k, HAC98, 0.5-min chase; lane l, HAC98, 1.0-min chase.