Difference between revisions of "Part:BBa K741018"

Latest revision as of 07:59, 3 October 2012

plac-RBS-cI-T-pRM-anticro-T-pCon(0.856)-RBS-crogfp-T

The fusion protein crogfp is consistently being produced. When the promoter plac is activated by IPTG, protein CI will be expressed to activate pRM. Then antisense RNA of cro will be expressed to inhibit the expression of CroGFP.This part is used to measure the function of pRM-anticro.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1589
Illegal NheI site found at 1612
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1370
Illegal BglII site found at 1706
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 2500

Figure 7. shows similar effects of glucose and IPTG on plac-cI-pRM-anticroT-pcon(0.856)-crogfp. 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-anticroT starts to work and antirco represses the translation of crogfp. On the contrast, glucose represses the expression of plac, which reduces the protein cI. pRM expresses less and the translation of crogfp is repressed less. Therefore, the unit fluorescence intensity of experimental groups with glucose added is the highest.

Data

plac-RBS-cI-T-pRM-anticro-T-pCon(0.856)-RBS-crogfp-T
OD
t/min	M9 medium	Parallel	0	500mM glucose	800mM glucose	1000mM glucose	1000mM IPTG
0	0.0559	1	0.1169	0.1229	0.1188	0.1245	0.1171
		2	0.1209	0.1194	0.1239	0.1132	0.1284
		3	0.1173	0.1191	0.1228	0.1163	0.113
45	0.0522	1	0.1733	0.1811	0.1819	0.1797	0.1682
		2	0.1756	0.1721	0.1808	0.1727	0.1739
		3	0.1723	0.1793	0.181	0.1793	0.1668
90	0.0543	1	0.2808	0.2779	0.2759	0.2876	0.253
		2	0.2768	0.2658	0.2708	0.2639	0.2489
		3	0.2795	0.2808	0.2605	0.286	0.2508
135	0.0541	1	0.3983	0.3923	0.3773	0.3943	0.3711
		2	0.3993	0.374	0.383	0.3816	0.3678
		3	0.3941	0.3969	0.4038	0.397	0.3787
180	0.0533	1	0.4662	0.4779	0.4944	0.4672	0.4641
		2	0.4747	0.4492	0.4634	0.4521	0.444
		3	0.4632	0.474	0.4483	0.4473	0.4509

fluorescence intensity
t/min	M9 medium	Parallel	0	500mM glucose	800mM glucose	1000mM glucose	1000mM IPTG
0	18.095	1	227.769	215.327	228.583	246.178	190.119
		2	234.019	229.854	236.364	216.548	221.432
		3	222.835	218.842	220.571	212.325	197.008
45	19.878	1	312.55	331.494	323.873	342.375	250.142
		2	294.057	338.868	326.819	333.868	252.672
		3	293.944	339.768	333.086	324.062	215.143
90	21.225	1	516.731	509.314	526.636	516.553	370.159
		2	499.503	500.773	467.699	501.155	353.996
		3	422.26	480.918	490.211	494.926	326.643
135	20.907	1	780.914	814.978	809.473	788.152	508.826
		2	788.354	793.653	764.974	780.294	494.73
		3	665.394	769.843	791.812	709.614	433.065
180	21.545	1	1057.396	1042.321	1055.53	1047.353	597.343
		2	1029.885	1073.57	1062.597	1012.626	570.092
		3	908.877	1084.629	1057.362	1026.325	527.67

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K741018 short</partinfo>
-The fusion protein crogfp downstream the constitutive promoter BBa_J23102 is continuously expressed. If IPTG exists, the cI will be expressed and then activates the pRM to produce the antisense RNA of cro(anticro). The anticro will repress the expression of crogfp.
+The fusion protein crogfp is consistently being produced. When the promoter plac is activated by IPTG, protein CI will be expressed to activate pRM. Then antisense RNA of cro will be expressed to inhibit the expression of CroGFP.This part is used to measure the function of pRM-anticro.
 <!-- Add more about the biology of this part here