Fig. 5.

Developing of E. coli O157 candidate conjugate in C. sedlakii MAGIC v.1. A schematic diagram of construction of C. sedlakii MAGIC v.1; B Coomassie stain of His-tagged CmeA protein purified from C.sedlakii and C. sedlakii MAGIC v.1 by nickel affinity chromatography. Biological samples were separated on a Bolt 4–12% bis–tris gel (Invitrogen) with MOPS buffer; C western blot analysis of CmeA purified from C. sedlakii and C. sedlakii MAGIC v.1, Biological samples were separated on a Bolt 4–12% bis–tris gel (Invitrogen) with MOPS buffer transferred to nitrocellulose membrane with an iBlot 2 dry blotting system. The membrane was probed with anti-His (Invitrogen) and anti-O157 (Abcam) antibody and detected with fluorescently labelled secondary antisera (green-His, red-O157) on a LI-COR Odyssey scanner. D glycosylation of CmeA in C. sedlakii MAGIC v.1 in broth and plate of His-tagged CmeA