Fig. 1.
Analysis of the C-terminal fragment rBacCPA279–363H6 with different consensus leader sequences. 7.5 × 106 Sf21 cells were infected in 10 ml of an EX-Cell 420 serum free medium with recombinant virus at a multiplicity of infection (MOI) of 2.0. After 72 h post-infection the infected cells were collected and lysed and the total protein of this preparation was quantified by the Bradford assay. Equal quantity of the cellular lysate of each sample was loaded electrophoresed on 12% SDS-PAGE gels and at the same time both (a) stained with Coomassie blue and (b) analysed in Western blot using an anti-His6-HRP conjugated MAb. Lane 1: rBac CPA279–363H6 without a leader sequence; lane 2: rBacCPA279–363H6 with L21 sequence; lane 3: rBacCPA279–363H6 with Kozaks sequence; lane 4: non-infected Sf21 cells (negative control); lane 5: Sf21 cell infected with recombinant baculovirus containing a target gene His tagged but not related with CPA (His6 positive control); lane 6: protein markers (the molecular weight bands in kDa, are reported on the right). The infections was repeated more than three times and the figure represents the highest-quality picture obtained