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. Author manuscript; available in PMC: 2011 Jun 2.
Published in final edited form as: Mol Microbiol. 2009 Dec 4;75(2):440–451. doi: 10.1111/j.1365-2958.2009.06993.x

Fig. 5.

Fig. 5

Secretion-competence of full-length OtpA in the periplasm.

A. HDB114 transformed with pPC20 (Ptrc-otpA) were incubated with IPTG and subjected to pulse-chase labelling. After cells were converted to a spheroplast suspension, half of the spheroplast suspension was untreated while the other half was treated with proteinase K (PK).

B. HDB114 transformed with pPC11 (PBAD-otpB) and pPC20 were incubated with IPTG and then subjected to pulse-chase labelling. Arabinose was added at the time of the chase. Cultures were divided into cell (C) and secreted (S) fractions, and the cell fraction was divided into three portions. One aliquot was untreated, the second was treated with PK, and the third was converted to a spheroplast suspension prior to PK treatment (sph). OtpA was immunoprecipitated from all samples. In part B, the S samples were derived from three times as much culture volume as each C and sph sample. m, molecular weight markers.