Figure 2

Bacterial expression of the modified ORFs of two alleles 1S^shx2.9 and 1Sshy2.3 in E. coli BL21 (DE3) and SDS-PAGE analysis of expressed products. The modified ORFs were prepared by removing the signal peptide sequence from each of the sequences by PCR mutagenesis. Protein extracts were prepared by dissolving cells directly in SDS-PAGE sample buffer. The glutenin proteins synthesized in E. coli directed by 1S^shx2.9 and 1Sshy2.3 under IPTG induction showed identical electrophoretic mobility to those from seeds of Ae. sharonensis (shown by arrows ). CK-x, y: proteins extracted from bacteria harbouring recombinant vectors pET–1S^shx2.9 or pET–1S^shy2.3 without IPTG induction for control; Aesh: proteins extracted from seeds of Ae. sharonensis.