Purification of recombinant GST-TAT-Apoptin
protein. SDS-PAGE(A) and Western-blot (B) analysis of the GST-TAT-Apoptinopt protein contained in various elution fractions collected from the GSTrap FF affinity column. The cytosolic extract of E. coli strain BL21(DE3) expressing GST-TAT-Apoptinopt protein was loaded onto a GSTrap FF column and the bound protein was eluted with elution buffer as described in Material and Methods. The eluted protein from the GSTrap FF affinity column was analyzed by SDS-PAGE and Western blotting using monoclonal anti-GST antibody. (C) Antigenicity analysis of GST-TAT-Apoptinopt. The purified GST-TAT-Apoptinopt was assayed by Western blotting using positive CAV-infected chicken serum. Lane M, pre-stained protein marker; lane 1, flow through; lane 2, fraction obtained after column washing, lane 3 and 4, eluted fraction 1 and 2, respectively, collecting after column elution.(D) Identity of the GST-opt-VP1 protein determined by MALDI-TOF. The bold letters represent actual amino acid matches to published amino acid sequence (Accession No. AF212490).