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. 2017 Aug 22;8:1597. doi: 10.3389/fmicb.2017.01597

FIGURE 2.

FIGURE 2

Archaeal TRAMs complemented the cold sensitivity of Escherichia coli BX04. (A) E. coli BX04 carries a quadruple deletion of cspA, B, G, and E genes, and its growth below 15°C is retarded. Cultures of E. coli strains overexpressing each of Mpsy_0643, Mpsy_2002, Mpsy_3043, Mpsy_3066, and cspE were adjusted to an optical density at 600 nm of 0.9 by dilution with fresh medium. Cultures were serially diluted 10-5-fold and spotted onto LB plates supplemented with ampicillin. The plates were incubated at 18, 22, and 37°C for 2–5 days with or without IPTG, as indicated (Supplementary Figure S4A). Growths at 37°C (10-5), 22°C (10-3), and 18°C (10-2) were shown. The pINIII cultures carry the empty plasmid vector. (B) Using the antibody against TRAM2002, a Western blot was performed to determine the abundance of four TRAM proteins in E. coli BX04. The empty plasmid pINIII and bacterial CspE were included as negative controls. The amount of loaded total protein for each TRAM gene cloned strain is shown beneath the gel. ± on the top of each lane indicates the presence or absence of IPTG.