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. 2010 Oct 25;79(1):279–287. doi: 10.1128/IAI.00821-10

FIG. 1.

FIG. 1.

(A) Purification of IbeA expressed in E. coli BL21(DE3). The whole ORF of ibeA was cloned into pET28a(+), leading to the expression of the 53.2-kDa fusion protein IbeA. BL21(DE3) cells were incubated in LB at 37°C with (lane 1) or without (lane 2) addition of IPTG, which induces the expression of IbeA. Protein of the total extracts (lanes 1 and 2) and the elution of the purified IbeA (lane 3) were separated on an SDS-PAGE gel and stained with Coomassie blue. Lane M, protein marker. (B) Immunoblotting analysis of total cell lysates prepared from DE205B, DE205M, DE205C, AAEC189P, and AAEC189C using a serum directed against IbeA. Expression of IbeA was detected in wild-type strain DE205B and complementation strains DE205C and AAEC189C but not the isogenic ibeA mutants. Lane M, prestained protein marker; lane 1, DE205B; lane 2, DE205M; lane 3, DE205C; lane 4, DE205P; lane 5, AAEC189P; lane 6, AAEC189C.