Induction of Boosted Immune Response in Mice by Leptospiral Surface Proteins Expressed in Fusion with DnaK
Figure 1
(a) Schematic representation of the expression cassettes. The genetically fused genes were obtained from individually cloned genes in pAE vector, and, then, surface protein genes were digested at PvuII/NcoI restriction sites and ligated in the same sites into pAE-DnaK construct. Depicted are T7 phage RNA polymerase promoter, ribosome-binding site (RBS), ATG start códon, 6X Histidine tag, restriction cloning sites, and the 2X (Gly-Pro) flexible hinge. (b) Analysis of purified recombinant proteins by SDS-PAGE. Purified recombinant protein eluted from Ni+2-charged Sepharose column with 1 M imidazole are visualized by Coomassie blue staining. Lane H (HMW) and L (LMW): high and low molecular mass protein markers; In kDa: lane 1: DnaK (68.9); lane 2: rLIC10494 (25.1); lane 3: DnaK-rLIC10494 (92.5); lane 4: rLIC12730 (75.7); lane 5: DnaK-rLIC12730 (143.1); lane 6: Lsa21 (19.8); lane 7: DnaK-Lsa21 (87.2); lane 8: Lp95 C-terminal (43.6); lane 9: DnaK-Lp95 C-terminal (110.9).
