Composite

Part:BBa_K741018

Designed by: Wuyang Chen   Group: iGEM12_USTC-China   (2012-08-29)
Revision as of 16:56, 30 September 2012 by Dunkle (Talk | contribs) (zs)

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

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.

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

USTC2012Result7 副本.png

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
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Parameters
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