Figure 4
From: A reinforced merging methodology for mapping unique peptide motifs in members of protein families
Epitope screening for mAb 3C1 and αRNase2. (A) The bacterial lysates containing RNase3-GFP (R31–133), RNase31–113-GFP (R31–113), RNase31–73-GFP (R31–73), RNase324–133-GFP (R324–133), RNase324–93-GFP (R324–93), RNase324–73-GFP (R324–73), RNase351–73-GFP (R351–73), RNase374–133-GFP (R374–133) and (B) RNase358–73-GFP (R358–73) were separated by 12% SDS/PAGE and separately probed by αHis and mAb 3C1. A fragment linking R31–133, R31–73, R324–93, R324–73, and R351–73 and GFP is 8.9 kD. (C) The schematic diagram of the chimeric constructions of RNase3/RNase2. The boxes labeled in grey, black and white indicate part of the $N$-terminus of RNase3, identical chimeric junctions and C-terminus of RNase2, respectively. Arg1 represents the first Arg residue in mature RNase3. (D) These chimeras were separated by 15{\%} SDS/PAGE (▶) and probed by αRNase2. (E) The bacterial lysates containing recombinant RNase258–73-GFP (R258–73, RNase273–90-GFP (R273–90) and RNase273–77-GFP (R273–77) were expressed and analyzed by Western blotting using $\alpha$ RNase2 and αHis.