Fig. 4.

A, Designing and testing MAGIC v.3. A nucleotide sequence of Biobricks promoters used in constructing MAGIC v.3 and their corresponding promoter strength [19]; B Western blot of 5 µg His-tagged CmeA protein purified by nickel affinity chromatography. Biological samples were separated on a Bolt 4–12% bis–tris gel (Invitrogen) with MOPS buffer and transferred to nitrocellulose membrane with an iBlot 2 dry blotting system. The membrane was probed with anti-His (Invitrogen) and anti- Ft-O antigen monoclonal antibody (Abcam) and detected with fluorescently labelled secondary antisera (green-His, red-Ft-O-antigen) on a LI-COR Odyssey scanner; H, densitometry analysis of glycoconjugate production in E. coli MAGIC v.3 Ft-O compared to E. coli bioconjugation Ft-O. Densitometry analysis of glycoconjugate was done from three biological replicates. Statistical analysis is from three biological replicates using Student’s t-test ns, p > 0.05; *,p < 0.05, **,p < 0.01, ***, p < 0.001