Difference between revisions of "Team:Nagahama/Results"

 
(71 intermediate revisions by 3 users not shown)
Line 1: Line 1:
 
{{Nagahama}}
 
{{Nagahama}}
−
[[File:Nagahama_POC実験.jpg|200px]]
+
=Result=
 +
 
 +
==Enhancement of farnesol resistance==
 +
 
 +
 
 +
[[File:MarA plate assay kamata did.png|750px|thumb|left|Fig. ?: Colony formation efficiencies of ''E. coli'' JM109 engineered with ''marA'' on geraniol overlaid plates.
 +
<br>
 +
''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|650px|thumb|center|Fig. 14: Comparison of colony numbers after addition of 0.5 %( v/v) geraniol hexane solution (geraniol solution).
 +
<br>
 +
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).]]
 +
 
 +
<br>
 +
<br>
 +
 
 +
 
 +
[[File:RT-PCR_Nagahama.png|400px|thumb|left|''Fig.The result of RT-PCR''
 +
<br>               
 +
Lane1:100bp DNA ladder 
 +
<br>
 +
Lane2:dxs(WT)
 +
<br>
 +
Lane3:dxs(MEP)
 +
<br>
 +
Lane4:idi(WT)
 +
<br>
 +
Lane5:idi(MEP)
 +
<br>
 +
Lane6:ispA(WT)
 +
<br>
 +
Lane7:ispA(MEP)
 +
<br>
 +
Lane8:ispD(WT)
 +
<br>
 +
Lane9:ispD(MEP)
 +
<br>
 +
Lane10:GES(WT)
 +
<br>
 +
Lane11:GES(GES)
 +
<br>
 +
Lane12:gapA(WT)
 +
<br>
 +
Lane13:gapA(MEP)
 +
<br>
 +
Lane14:gapA(GES)
 +
]]
 +
[[File:RT-PCR Negative control_Nagahama.png|370px|thumb|none|''Fig.The result of RT-PCR Negative control''
 +
<br>
 +
Lane1:dxs(WT)
 +
<br>
 +
Lane2:dxs(MEP)
 +
<br>
 +
Lane3:idi(WT)
 +
<br>
 +
Lane4:idi(MEP)
 +
<br>
 +
Lane5:ispA(WT)
 +
<br>
 +
Lane6:ispA(MEP)
 +
<br>
 +
Lane7:ispD(WT)
 +
<br>
 +
Lane8:ispD(MEP)
 +
<br>
 +
Lane9:GES(WT)
 +
<br>
 +
Lane10:GES(GES)
 +
<br>
 +
Lane11:gapA(WT)
 +
<br>
 +
Lane12:gapA(MEP)
 +
<br>
 +
Lane13:gapA(GES)
 +
]]
 +
 
 +
 
 +
We reinforced by overexpressing genes that dxs,idi,ispD and ispF, which was the bottom neck of MEP route. GC-MS detected synthetic amount of farnesol was very low. We checked amount of transcription by RT-PCR and q-PCR.
 +
 
 +
 
 +
<br />
 +
<br />
 +
<br /><br /><br /><br />
 +
 
 +
 
 +
 
 +
 
 +
 
 +
 
 +
==temp==
 +
 
 +
Here you can describe the results of your project and your future plans.
 +
What should this page contain?
 +
Clearly and objectively describe the results of your work.
 +
Future plans for the project
 +
Considerations for replicating the experiments
 +
Project Achievements
 +
You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.
 +
A list of linked bullet points of the successful results during your project
 +
A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.
 +
Inspiration
 +
See how other teams presented their results.
 +
2014 TU Darmstadt
 +
2014 Imperial
 +
2014 Paris Bettencourt

Latest revision as of 01:46, 2 October 2016

MENU ▤
　　

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)


We reinforced by overexpressing genes that dxs,idi,ispD and ispF, which was the bottom neck of MEP route. GC-MS detected synthetic amount of farnesol was very low. We checked amount of transcription by RT-PCR and q-PCR.











temp

Here you can describe the results of your project and your future plans. What should this page contain? Clearly and objectively describe the results of your work. Future plans for the project Considerations for replicating the experiments Project Achievements You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer. A list of linked bullet points of the successful results during your project A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort. Inspiration See how other teams presented their results. 2014 TU Darmstadt 2014 Imperial 2014 Paris Bettencourt