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. 2014 May 9;9(5):e96757. doi: 10.1371/journal.pone.0096757

Figure 3. Purification of the wild type and N-terminal deletion BirA proteins and the biotin acceptor proteins.

Figure 3

The proteins were purified as described in Materials and Methods and subjected to SDS-electrophoresis on a 4–20% polyacrylamide gel. M: molecular weight standards (Precision Plus Protein Standard Kaleidoscope from BioRad). Lane 1: B. subtilis N-terminally hexahistidine-tagged BirA (38.9 kDa). Lanes 2-5. B. subtilis N-terminally hexahistidine-tagged Δ2-63 BirA (31.8 kDa), Δ2-65 BirA (31.6 kDa), Δ2-74 BirA (30.4 kDa) and Δ1-81 BirA (29.7 kDa), respectively. Lanes 6-11 are the B. subtilis acceptor proteins AccB-86 (9.4 kDa), PyC-77 (8.3 kDa) and biotin lipoyl attachment protein (BLAP) (8.73 kDa). Lane 10 is E. coli C-terminal hexahistidine-tagged BirA. Lane 11 is E. coli C-terminal hexahistidine tagged Δ2-65 BirA (29.18 kDa) and lane 11 is E. coli AccB-87.