Figure 1
Design, expression, purification and characterization of the baculovirus-expressed recombinant human ZP1 273-551aa . Panel (a) represents the schematic diagram of the cDNA encoding human ZP1273-551aa with 6× His tag (His6) at C-terminus cloned in baculovirus transfer vector, pAcGP67-A, using BamH I and EcoR I restriction sites downstream of gp67 insect secretory signal sequence under polyhedrin promoter (Ppol). Sf 21 cells (0.8 × 106) were infected with the recombinant or wild type AcNPV virus. After 96 h, the infected cells and supernatant were harvested and processed for Western blot as described in Methods. Panels (b) & (c) represent immunoblot showing the expression of ZP1273-551aa in cell pellet and 10× concentrated culture supernatant respectively from Sf 21 cell infected with ZP1273-551aa-recombinant virus. Lanes 1 and 2 represent the cell pellet or culture supernatant of Sf 21 cell infected with wild type AcNPV and ZP1273-551aa-recombinant virus respectively. Panel (d) represents Coomassie stained SDS-PAGE profile of ZP1273-551aa purified from the cell pellet (lane 1, 5 μg/lane). Lane M represents molecular weight markers.