Part:BBa_K741004
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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 | Parellel | 0 | 500mM gl | 800mM gl | 1000mM g | 1000mM I |
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 | ||
plac-RBS-cI-T-pRM-RBS-GFP-T | |||||||
fluorescence intensity | |||||||
t/min | M9 medium | Parellel | 0 | 500mM gl | 800mM gl | 1000mM g | 1000mM I |
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 |
None |