Cofactor Independent Phosphoglycerate Mutase of Brugia malayi Induces a Mixed Th1/Th2 Type Immune Response and Inhibits Larval Development in the Host
Figure 2
Cloning, expression, and purification of Bm-iPGM. (a) Purification of Bm-iPGM: L1: flow through; L2-L4: wash 1–3; L5 and L6: elute 1-2; L7: standard protein marker (kDa). (b) Western blot analysis using anti-His mAb: L1: prestained protein marker; L2: purified Bm-iPGM. (c) MALDI-TOF analysis of the molecular mass of recombinant Bm-iPGM. A single major peak confirmed the mass of recombinant Bm-iPGM to be 61.799 kDa. (d) Far-UV CD spectra of Bm-iPGM; CD measurements were made on JASCO J810 spectropolarimeter calibrated with ammonium (+)-10-camphorsulfonate with 6 μM protein in 10 mM CGH buffer. (e) Fluorescence emission spectra of Bm-iPGM and spectra of Bm-iPGM in 50 mM phosphate buffer were recorded with Perkin Elmer LS50B luminescence spectrometer. On excitation at 280 nm, maximum emission spectra were noted at 340 nm.
