Target bimA gene was cloned in a pET28a expression
vector, and transformation was carried out in Escherichia
coli BL21 (DE3) cells for expression of recombinant BimA
protein and one clone (henceforth called pETBimA), which
showed maximum expression after 4 hours of induction by
0.5-mM isopropyl-b-D-1-thiogalactopyranoside (IPTG).
Expressed protein was obtained in insoluble fraction (inclusion
bodies) and puried by immobilized metal affinity
chromatography (IMAC) using a Ni4-NTA fast-flow
chelating Sepharose column. The purried protein was
dialyzed against 0.01-M phosphate-buffered saline (PBS;
pH 7.2) containing decreasing concentrations of urea as
reported previously