Team:Nagahama/Results

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Result

Enhancement of farnesol resistance

Fig. ?: Colony formation efficiencies of E. coli JM109 engineered with marA on geraniol overlaid plates.
E. coli JM109 and E. coli JM109 (marA) were spotted on LBGMg agar plates in serial ten-fold dilutions (10⁻¹～10⁻6), overlaid with 30.0 % (v/v)　fanesol hexane solution (farnesol solution), and incubated at 30°C for 24 h. This figure shows that E. coli JM109 (marA) cells that overexpress the marA product is more survived on 30.0 % farnesol solution overlay plates than the counterpart control E. coli JM109 wild type cells.



File:MarA plate assay nishikawa last 2.png
Fig. 14: Comparison of colony numbers after addition of 0.5 %( v/v) geraniol hexane solution (geraniol solution).
Time interval for treatment was set every 1 hour from 1 hour to 4 hours. A: E. coli JM109 (WT) + hexane; B: E. coli JM109 (marA) + hexane; C: E. coli JM109 (WT) + 0.5 % geraniol solution; D: E. coli JM109 (marA) + 0.5 % geraniol solution. As shown in Figs. 14 A and B, treatment with hexane of E. coli JM109 (WT) and of E. coli JM109 (marA) showed similar colony numbers during these treatment intervals to those of time zero. This result suggests that hexane at this concentration and duration of time for 4hours did not affect both cell growth. In contrast, treatment with geraniol of E. coli JM109 (WT) and of E. coli JM109 (marA) showed toxicities to both strains (Figs. 14 B, C and D). If we watch the colony numbers carefully, E. coli JM109 (marA) had more than E. coli JM109 (WT) during these treatment intervals ((Figs. 14 C and D). These results demonstrate that toxicity of the geraniol was less to the strain E. coli JM109 (marA) than the strain E. coli JM109 (WT).




Fig.The result of RT-PCR
Lane1:100bp DNA ladder
Lane2:dxs(WT)
Lane3:dxs(MEP)
Lane4:idi(WT)
Lane5:idi(MEP)
Lane6:ispA(WT)
Lane7:ispA(MEP)
Lane8:ispD(WT)
Lane9:ispD(MEP)
Lane10:GES(WT)
Lane11:GES(GES)
Lane12:gapA(WT)
Lane13:gapA(MEP)
Lane14:gapA(GES)


Fig.The result of RT-PCR Negative control
Lane1:dxs(WT)
Lane2:dxs(MEP)
Lane3:idi(WT)
Lane4:idi(MEP)
Lane5:ispA(WT)
Lane6:ispA(MEP)
Lane7:ispD(WT)
Lane8:ispD(MEP)
Lane9:GES(WT)
Lane10:GES(GES)
Lane11:gapA(WT)
Lane12:gapA(MEP)
Lane13:gapA(GES)
||<#ffffff' style='text-align:center;width:525px' ``
Fig.The result of RT-PCR
Lane1:100bp DNA ladder
Lane2:dxs(WT)
Lane3:dxs(MEP)
Lane4:idi(WT)
Lane5:idi(MEP)
Lane6:ispA(WT)
Lane7:ispA(MEP)
Lane8:ispD(WT)
Lane9:ispD(MEP)
Lane10:GES(WT)
Lane11:GES(GES)
Lane12:gapA(WT)
Lane13:gapA(MEP)
Lane14:gapA(GES)
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