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Team:KU Seoul/Result
From 2009.igem.org
Revision as of 14:09, 20 October 2009 by
Cheolwon.choi
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Home
Team
Project
Overview
Materials
Experiment Diary
Result
Parts
Notebook
Safety
Sponsors
Results
1. Synthetic Circuit - I : pSB3C5/Pars-gfp-Pznt-rfp
1)Pznt-rfp
This parts is producing red fluorosence based on the zinc ion level. The zinc detecting promoter can be
induced beyond 1mM concentration of zinc ion. This range from 1mM to 2mM is proper to detect zinc level on
the common waste water. Maybe it is required to detect the upper concentration of zinc ion in waste water that
using the dilution method which dilute the level of zinc concentration. If the level curve is too rugh or not
too much change according to concentraion, you need to apply dilution or concentration of sample.
Zn
2+
(mM)
0
0.5
0.75
1
1.5
2
OD
600
0.490 ± 0.003
0.459 ± 0.004
0.433 ± 0.008
0.418 ± 0.005
0.275 ± 0.011
0.243 ± 0.008
Red fluorescence (RF)
26628 ± 706
27856 ± 292
29333 ± 292
31217 ± 66
46044 ± 237
49188 ± 174
OD
600
/RF
54376 ± 1203
60646 ± 787
67803 ± 573
74687 ± 755
167833 ± 7202
202565 ± 6428
Background
0
6270 ± 520
13427 ± 1719
20311 ± 475
113457 ± 6016
148189 ± 6731
2)Pars-gfp
Differing from zinc ion, Arsenic ion is more severe impact on the E.coli. When microbials are exposed by
arsenic ion even 1mM concentration, This concentraion of arsenic ion is already full of toxicity. It is very
senstive to detect arsenic ion. It means that arsenic ion can provide strong harmful effect on the organism
even microbial which is known as the best survivor on the earth. The interaction of arsenic ion is participated
in more than two or more group of proteins or others. See the curve of fluorosence of GFP.the level curve is
show sigmoid curve form.
It is supose that the the cost of the expression of arsenic ion binder is very expensive for microbial.
Because even lower level of concentraion the fluorosence curve did not show regression curve at the initiation
point. Strong inhibitor can be exist at the initation of expression state.
AsO
3-
(mM)
0
0.00625
0.125
0.25
0.5
0.75
1
OD
600
0.553 ± 0.009
0.519 ± 0.002
0.480 ± 0.005
0.395 ± 0.007
0.315 ± 0.006
0.286 ± 0.004
0.273 ± 0.006
Green fluorescence (GF)
27740 ± 226
26411 ± 169
25267 ± 267
25536 ± 213
25407 ± 259
24849 ± 152
26479 ± 149
OD
600
/GF
50206 ± 1225
50855 ± 123
52606 ± 689
64669 ± 1684
80665 ± 603
86897 ± 1321
96784 ± 2165
Background
0
648 ± 1314
2399 ± 969
14463 ± 2896
30459 ± 1631
36691 ± 1636
46578 ± 3220
2.Synthetic circuit – II : PyodA-AMD
Cd
2+
(mM)
0
0.05
0.1
0.2
0.4
0.8
OD
615
0.242 ± 0.005
0.258 ± 0.008
0.274 ± 0.011
0.284 ± 0.01
0.293 ± 0.011
0.302 ± 0.01
Background
0
0.016 ± 0.004
0.032 ± 0.006
0.043 ± 0.005
0.052 ± 0.007
0.060 ± 0.007