Figure 1. Construction, expression and purification of rec-pediocin.
a) Deduced amino acid sequences of the recombinant pre-pediocin. Authentic portion of the sequences are shown in bold. The added tags are in italics and changed nucleotide primers generated aa in bold italics.
b) SDS-PAGE (15% acrylamide) gel analysis of IBs purified from hyper expressing E. coli. Lane 1, whole cell lysate from induced cells. Lane 2, 3 & 4 are IBs washed with 2, 4 and 6 M urea. M, is medium range protein marker (Bangalore Genei). Arrow indicates the position of 12.8 kDa rec-pediocin.
c) Single step purification of rec-pediocin. Gel analysis of 6 X histidine -Xpress-prepediocin produced in E. coli. Lane 1, total cell lysate uninduced; 2, Lysate, induced recombinant and lane 3, Ni-NTA beads absorbed IBs protein eluted with 0.5 M immidazole. M, protein marker. Arrow indicates purified rec-pediocin.