Fig 1: Molecular cloning of chiA-3 and expression of ChiA-3-6xHis. (a) InstantBlue-stained acrylamide gel of proteins present in supernatants and cell pellet lysates from cultures grown at 23 °C supplemented with arabinose (induction +) or glucose (induction -). No induced bands were easily discerned. (b) Western immunoblot produced from an identically loaded acrylamide gel to that presented in (a) run in parallel with the gel in (a) and probed with an a-6xHis antibody (see Methods). A band corresponding to the expected molecular weight of ChiA-3-6xHis (48.51 kDa) was only detected in the cell pellet lysate of E. coli harbouring pMJD157 (plasmid +). This size is consistent with the retention of the fusion protein without the cleavage of the putative signal sequence. Protein ladders: NEB #P7719S and #P7717S. EV=empty vector (pBAD33).