Difference between revisions of "Part:BBa K741004"

Revision as of 00:33, 3 October 2012

plac-RBS-cI-T-pRM-RBS-GFP-T

Protein CI is expressed to activated the promoter pRM. Thus different concentration of CI lead to different strength of green fluorescence. We use this part to test the character of modified pRM being activated by CI.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1898

Figure 3 shows similar effects of glucose and IPTG on plac-cI-pRM-gfp. pRM is activated by the protein cI. By adding IPTG, plac is activated, which leads to the production of the protein cI. With the existence of cI, pRM-GFP starts to work. On the contrast, glucose represses the expression of plac, which reduces the protein cI. Therefore, pRM expresses little and the unit fluorescence intensity of experimental groups with glucose added is the lowest.

Figure 4 shows the difference of the unit fluorescence intensity between plac-gfp and plac-cI-pRM-gfp. Generally, the fluorescence intensity of plac-gfp is higher than that of plac-cI-pRM-gfp, because the transcription efficiency of pRM is quite low, which reduces the expression of gfp in comparison with plac-gfp.

Data

plac-RBS-cI-T-pRM-RBS-GFP-T
OD
t/min	M9 medium	Parallel	0	500mM glucose	800mM glucose	1000mM glucose	1000mM IPTG
0	0.0559	1	0.1157	0.1178	0.117	0.1167	0.1119
		2	0.1086	0.1141	0.1142	0.1113	0.1111
		3	0.1177	0.1222	0.1252	0.1074	0.1056
45	0.0522	1	0.1597	0.1697	0.1661	0.1684	0.1515
		2	0.152	0.163	0.1682	0.162	0.1529
		3	0.1564	0.1702	0.1777	0.1575	0.1455
90	0.0543	1	0.2221	0.2305	0.238	0.2329	0.2082
		2	0.2072	0.2171	0.224	0.2306	0.2082
		3	0.219	0.2267	0.2449	0.2202	0.1643
135	0.0541	1	0.3091	0.3388	0.3365	0.3298	0.3063
		2	0.2947	0.3177	0.334	0.3291	0.3003
		3	0.3221	0.3278	0.3265	0.3186	0.2989
180	0.0533	1	0.4281	0.4068	0.3955	0.4129	0.3895
		2	0.3772	0.4165	0.4418	0.4285	0.3804
		3	0.4002	0.4422	0.4643	0.3929	0.3789


fluorescence intensity
t/min	M9 medium	Parallel	0	500mM glucose	800mM glucose	1000mM glucose	1000mM IPTG
0	18.095	1	356.895	353.702	342.454	289.758	326.681
		2	364.212	341.485	344.079	332.975	338.72
		3	333.663	329.681	334.645	330.905	327.423
45	19.878	1	428.805	440.026	433.171	429.13	421.519
		2	433.472	428.521	434.422	434.594	429.878
		3	383.44	430.936	395.792	431.248	431.64
90	21.225	1	582.263	522.888	514.303	519.709	553.375
		2	502.785	531.559	524.747	517.783	552.984
		3	482.716	504.875	505.268	506.006	525.12
135	20.907	1	697.398	632.717	615.29	589.652	700.113
		2	652.66	585.696	597.532	580.199	664.366
		3	604.926	621.743	607.306	581.586	625.487
180	21.545	1	714.418	719.636	694.03	638.407	717.069
		2	721.891	686.106	641.981	625.515	844.074
		3	730.621	660.123	654.52	555.481	628.488

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K741004 short</partinfo>
-Protein CI is expressed to activated the promoter pRM. Thus different concentration of CI lead to different strength of green fluorescence.
+Protein CI is expressed to activated the promoter pRM. Thus different concentration of CI lead to different strength of green fluorescence. We use this part to test the character of modified pRM being activated by CI.
 <!-- Add more about the biology of this part here