Figure 3.

Purification of recombinant cotton annexin. The cotton annexin cDNA was expressed in E. coli BL21 (DE3) as His-tagged protein and purified by affinity chromatography as described previously. Proteins were separated by 10% SDS-PAGE and stained with Coomassie blue. Lane 1, Crude extract from uninduced cell; lane 2, crude extract from isopropyl 1-thiol-β-d-galactopyranoside-induced cell; lane 3, soluble fraction obtained from lane 2 fraction; lane 4, washed-out fraction from His-tag chromatography; and lane 5, eluted fraction from His-tag chromatography. Note the highly purified annexin band in lane 5 with an apparent molecular mass of 36 kD.