Figure 6
Cell-wall binding efficiency analysis of recombinant B. thuringiensis cells anchored by various Mbg domains. (A) Western blot analysis of cell-wall fractions. All fractionated samples were prepared in parallel, and an equal volume of each sample was used for analysis. The relative amount of fusion protein shown on each lane was quantified using the amount of (Mbgn)2-GFP as the reference value (100%). Abbreviations: Mbgn, the N-terminal domain of Mbg; Mbgc, the C-terminal domain of Mbg; lysM, the lysin motif; GFP, green fluorescence protein; (Mbgn)2, two tandemly aligned Mbgn repeats; (Mbgn)3, three tandemly aligned Mbgn repeats. (B) Total GFP fluorescence intensities of BMB171 and the recombinant B. thuringiensis cells expressing GFP-fusion proteins with various anchoring domains. (C) Flow cytometric analysis of BMB171 and the recombinant B. thuringiensis cells expressing GFP-fusion proteins with various anchoring domains. (a) BMB171; (b) MB163 cells expressing Mbg-GFP; (c) MB164 cells expressing Mbgn-GFP; (d) MB166 cells expressing LysM1-GFP; (e) MB168 cells expressing (Mbgn)2-GFP; and (f) MB169 cells expressing (Mbgn)3-GFP. The value of each histogram indicates the percentage of total Cy5-labelled fluorescent cells. For each experiment, 100,000 cells were analyzed.