Difference between revisions of "Part:BBa K3081053"
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Quorum sensing system is widely used in gene circuit design as a sensor of cell density. As a transcription factor, LuxR senses AHL and then activate the transcript of plux. In our project, we want dCas9 to express when bacteria density is high. So we put the dCas9 gene under the control of plux, and get this composite part. And we can use golden-gate assembly methods to change the sequence of sgRNA. | Quorum sensing system is widely used in gene circuit design as a sensor of cell density. As a transcription factor, LuxR senses AHL and then activate the transcript of plux. In our project, we want dCas9 to express when bacteria density is high. So we put the dCas9 gene under the control of plux, and get this composite part. And we can use golden-gate assembly methods to change the sequence of sgRNA. | ||
| − | https://2019.igem.org/wiki/images/b/b5/T--Peking--QS3-.png | + | <center>https://2019.igem.org/wiki/images/b/b5/T--Peking--QS3-.png</center> |
| − | Figure1 We transfer this part in <i>E. coli</i> ,BL21(DE3), and culture them in M9 medium containing a series of AHL concentration for 6 hours. Receiver cells show remarkable dose-dependent change in length with addition of AHL. | + | <center>Figure1 We transfer this part in <i>E. coli</i> ,BL21(DE3), and culture them in M9 medium containing a series of AHL concentration for 6 hours. Receiver cells show remarkable dose-dependent change in length with addition of AHL. </center> |
| − | https://2019.igem.org/wiki/images/7/75/T--Peking--QS6-.png | + | <center>https://2019.igem.org/wiki/images/7/75/T--Peking--QS6-.png</center> |
| − | Figure2 The Pattern diagram of Quorum sensing system. | + | <center>Figure2 The Pattern diagram of Quorum sensing system.</center> |
| − | https://2019.igem.org/wiki/images/0/01/T--Peking--QS7.png | + | <center>https://2019.igem.org/wiki/images/0/01/T--Peking--QS7.png</center> |
| − | https://2019.igem.org/wiki/images/d/de/T--Peking--QS88-.png | + | <center>https://2019.igem.org/wiki/images/d/de/T--Peking--QS88-.png</center> |
Figure3 We transferred this part into <i>E. coli</i>, BL21(DE3) as receptor cells, and drop or coat them on the solid medium like A. Then drop 5 μl M9 containing 0.5 Mm IPTG. Solid medium were incubated about 16h. The bacteria density in one drop showed great difference with different distances from IPTG. | Figure3 We transferred this part into <i>E. coli</i>, BL21(DE3) as receptor cells, and drop or coat them on the solid medium like A. Then drop 5 μl M9 containing 0.5 Mm IPTG. Solid medium were incubated about 16h. The bacteria density in one drop showed great difference with different distances from IPTG. | ||
Revision as of 02:37, 21 October 2019
AHL receiver device to expression of dCas9 and sgRNA
Quorum sensing system is widely used in gene circuit design as a sensor of cell density. As a transcription factor, LuxR senses AHL and then activate the transcript of plux. In our project, we want dCas9 to express when bacteria density is high. So we put the dCas9 gene under the control of plux, and get this composite part. And we can use golden-gate assembly methods to change the sequence of sgRNA.
Figure3 We transferred this part into E. coli, BL21(DE3) as receptor cells, and drop or coat them on the solid medium like A. Then drop 5 μl M9 containing 0.5 Mm IPTG. Solid medium were incubated about 16h. The bacteria density in one drop showed great difference with different distances from IPTG.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1321
Illegal XhoI site found at 5538 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5728
Illegal BsaI.rc site found at 1004
Illegal BsaI.rc site found at 5324