e102 Nucleic Acids Research, 2008, Vol. 36, No. 16 PAGE 6 OF 10 Figure 2. Various phenotypic changes induced by ATFs. (A) A thermotolerant phenotype induced by ATFs. The growth of thermotolerant cells (T1, 2 and 3) and control cells (C) was monitored after incubation for 2 h at 378C (No shock) or for 2 h at 558C (Heat shock). The triangles above each panel indicate tenfold serial dilutions (1 : 1 to 1 : 1000, left to right) of spotted cells. (B) An osmotolerant phenotype induced by ATFs. The growth of cells was monitored on minimal A plates containing 0.6 M NaCl. The triangles above each panel indicate ﬁvefold serial dilutions (1 : 1 to 1 : 25, left to right) of spotted cells. OT1-OT4, osmotolerant cell isolates 1–4, respectively. (C) A cold-tolerant phenotype induced by ATFs. Cells were cultured at 158C, and cell growth was monitored. The triangles above each panel indicate ﬁvefold serial dilutions (1 : 1 to 1 : 125, left to right) of spotted cells. CT1 and CT2, cold-tolerant cell isolates 1 and 2, respectively. For all panels, C indicates the wild-type E. coli transformed with a control plasmid without any previous selection. After the ﬁrst round of phenotypic screening, all of the phenotypic changes were conﬁrmed by plasmid rescue, sequence analysis and retransformation.