Confirmation of Lpp-OmpA-multimeric Buf IIIb-L fusion genes and the expression of LO-B_n fusion proteins.

<p>(A) The number of Buf IIIb-L genes cloned in pLpp-OmpA-B_n was determined by digesting with <i>Not</i>I, whose sites flank each multimer. Lanes 1–4 represent <i>Not</i>I-digested pLpp-OmpA-B₀, -B₁, -B₂, and -B₃, which contains 0, 1, 2, or 3 copies of the Buf IIIb-L gene, respectively. Lane M represents size markers. (B) Total cell proteins were analyzed using SDS-PAGE. Lanes 1–4 represent total cell proteins from <i>E. coli</i> BL21 (DE3) harboring pLO-B₀, -B₁, -B₂, and -B₃, respectively. Lane M represents molecular weight markers. The <i>closed triangles</i> indicate the LO-B_n fusion proteins expressed from each clone. (C) The presence of LO-B_n fusion proteins in the total cell proteins from each clone was confirmed by Western blot using anti-His antibody. The bands corresponding to each LO-B_n fusion proteins (n = 0, 1, 2, and 3) were detected in lanes 1–4 (13.1, 15.85, 18.60, and 21.35 kDa, respectively).