The plasmid map and a schematic representation (A), the SDS-PAGE (B) and the western blotting analysis (C) of the Protein A-CauloGA fusion protein. (A) The plasmid map of pEZZ18-CauloGA (upper) and a schematic representation of the Protein A-CauloGA fusion protein (lower). The construction of the plasmid is detailed in Materials and Methods. P lac, Protein A and Amp r denote the lac promoter, Protein A and the ampicillin-resistant genes of the pEZZ18 vector, respectively. The gray arrow represents the CauloGA gene. The dark and light shaded boxes represent Protein A (synthetic ZZ domain) and CauloGA, respectively. N and C indicate the N- and C- termini of the protein. (B) The SDS-PAGE analysis of the crude cell extract and the IgG Sepharose-purified CauloGA. Lane M, molecular weight marker (Precision Plus Protein™ Standards, Bio-Rad laboratories); lane 1, crude extract, supernatant from cell homogenates; and lane 2, purified CauloGA eluted from IgG Sepharose. The numbers in the margin represent the molecular masses (kDa) of the proteins in the molecular weight marker. (C) The Western blotting analysis of the purified CauloGA. The numbers represent the molecular masses (kDa) of the proteins in the molecular weight marker.