Measurement

Part:BBa_K741004

Designed by: Wuyang Chen   Group: iGEM12_USTC-China   (2012-07-24)

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 (BBa_I12007) 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

USTC2012Result3 副本.png

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.

USTC2012Result4 副本.png

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
[edit]
Categories
Parameters
None