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. 2018 Oct 10;179(3):958–968. doi: 10.1104/pp.18.01085

Figure 2.

Figure 2.

Tests of functional complementation of an E. coli ΔthiG strain by diverse bacterial THI4s. A, The ΔthiG strain harboring the pBAD24 vector alone or containing a THI4 gene was cultured in MOPS minimal medium containing 0.2% (w/v) glycerol and 0.02% (w/v) arabinose, plus or minus 1 mm Cys, in aerobic or anaerobic conditions. Aerobic controls supplemented with 100 nm thiamin were included. Anaerobic media contained 40 mm nitrate as the electron acceptor. Overnight liquid cultures of three independent clones for each construct were 10-fold serially diluted and spotted on the plates. Images were captured after incubation at 37°C for 7 d. B, Growth in liquid MOPS medium as above, plus 1 mm Cys, of wild type E. coli (WT) and the ΔthiG strain harboring pBAD24 alone (EV) or containing T. ammonificans THI4 (TaTHI4). Cultures were incubated in aerobic conditions at 37°C. C, Thiamin monophosphate (ThMP) and diphosphate (ThDP) contents of wild type and TaTHI4-complemented cells from B, harvested when OD600 reached 1.0–1.5; free thiamin was undetectable. Values in B and C are means and SE for three independent replicates. The difference in ThDP content between wild type and TaTHI4-complemented cells was significant at P < 0.001 (**), as determined by Student’s t test. Where no error bars appear in B they are smaller than the symbol.