Difference between revisions of "Part:BBa F2620"
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'''Description'''<br> | '''Description'''<br> | ||
| − | A transcription factor, LuxR (''<partinfo>C0062</partinfo>'') that is active in the presence of cell-cell signaling molecule [[3OC6HSL|3OC<sub>6</sub>HSL]] is controlled by a TetR (''<partinfo>BBa_C0040</partinfo>'') regulated operator (''<partinfo>BBa_R0040</partinfo>''). ''Device input'' is [[3OC6HSL|3OC<sub>6</sub>HSL]]. ''Device output'' is [[PoPS]] from a LuxR-regulated operator. If used in a cell containing TetR then a second input signal such as [http://openwetware.org/wiki/ATc aTc] can be used to produce a logical AND function. Get the pdf datasheet [[:Image: | + | A transcription factor, LuxR (''<partinfo>C0062</partinfo>'') that is active in the presence of cell-cell signaling molecule [[3OC6HSL|3OC<sub>6</sub>HSL]] is controlled by a TetR (''<partinfo>BBa_C0040</partinfo>'') regulated operator (''<partinfo>BBa_R0040</partinfo>''). ''Device input'' is [[3OC6HSL|3OC<sub>6</sub>HSL]]. ''Device output'' is [[PoPS]] from a LuxR-regulated operator. If used in a cell containing TetR then a second input signal such as [http://openwetware.org/wiki/ATc aTc] can be used to produce a logical AND function. Get the pdf datasheet [[:Image:EndyFig1-F2620DataSheet.pdf|here]]. |
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|Not measured | |Not measured | ||
|- | |- | ||
| − | |rowspan="2"|[[Transcription Demand|''Transcriptional output demand:'']]<br>(Low/High Input) | + | |rowspan="2"|[[Transcription Demand|''Transcriptional output demand:'']]<br>(Low/High Input)<br>Nt = length of downstream transcript in nucleotides |
| − | |( | + | |(0/6xNt) nucleotides cell<sup>-1</sup> s<sup>-1</sup> |
|- | |- | ||
| − | |( | + | |(0/1.5E-1xNt) RNAP cell<sup>-1</sup> |
|- | |- | ||
|[[Growth Rate|''Growth Rate'']]<br>(Low/High Input) | |[[Growth Rate|''Growth Rate'']]<br>(Low/High Input) | ||
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'''Compatibility'''<br> | '''Compatibility'''<br> | ||
[https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=cell ''Chassis:''] Device has been shown to work in ''<partinfo>BBa_V1000</partinfo>'',''<partinfo>BBa_V1001</partinfo>'',''<partinfo>BBa_V1002</partinfo>'', [[Chassis/Cell-Free_Systems/Commercial_E.coli_S30| E.Coli S30 Extract]] [[Chassis/Cell-Free_Systems/Commercial_E.coli_S30/F2620 | (data)]]<br> | [https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=cell ''Chassis:''] Device has been shown to work in ''<partinfo>BBa_V1000</partinfo>'',''<partinfo>BBa_V1001</partinfo>'',''<partinfo>BBa_V1002</partinfo>'', [[Chassis/Cell-Free_Systems/Commercial_E.coli_S30| E.Coli S30 Extract]] [[Chassis/Cell-Free_Systems/Commercial_E.coli_S30/F2620 | (data)]]<br> | ||
| − | [[ | + | [[Plasmid backbones|''Plasmids:'']] Device has been shown to work on ''<partinfo>pSB3k3</partinfo>'' and ''<partinfo>pSB1A3</partinfo>''<br> |
[[Part Types|''Devices:'']] Device has been shown to work with ''<partinfo>BBa_E0430</partinfo>'', ''<partinfo>BBa_E0434</partinfo>'', ''<partinfo>BBa_E0240</partinfo>''<br> | [[Part Types|''Devices:'']] Device has been shown to work with ''<partinfo>BBa_E0430</partinfo>'', ''<partinfo>BBa_E0434</partinfo>'', ''<partinfo>BBa_E0240</partinfo>''<br> | ||
Crosstalk with systems containing ''<partinfo>BBa_C0040</partinfo>''.<br> | Crosstalk with systems containing ''<partinfo>BBa_C0040</partinfo>''.<br> | ||
[[Help:Signalling|''Cell-Cell Signaling Systems:'']] Crosstalk with devices using [[3OC6HSL|3OC<sub>6</sub>HSL]], C6HSL, C7HSL, C8HSL, C10HSL. | [[Help:Signalling|''Cell-Cell Signaling Systems:'']] Crosstalk with devices using [[3OC6HSL|3OC<sub>6</sub>HSL]], C6HSL, C7HSL, C8HSL, C10HSL. | ||
</div> | </div> | ||
| + | '''Contribution: Arizona_State 2016''' | ||
| + | |||
| + | Authors: Ernesto Luna, Brady Dennison, Cassandra Barrett, Jimmy Xu, Jiaqi Wu, Dr. Karmella Haynes | ||
| + | |||
| + | The 2016 Arizona_State team ran a series of induction tests on 10 AHL Senders (naturally-produced by an BL21 E.coli chassis) and 5 synthetic AHL variants using Bba_F2620 as the output device. This included a series of visual plate induction tests as well as an 8-hour GFP absorbance read. This contribution aims to expand the characterization of this part by identifying crosstalk partners as well as potential orthogonal systems. | ||
| + | |||
| + | The table below summarized the findings of our induction test. Note: these results are not conclusive and are simply what the collected data indicated from our 8-hour induction test. | ||
| + | |||
| + | |||
| + | |||
| + | Two runs at two different concentrations (100nM and 1uM) using synthetic AHLs yielded some interesting differences, with synthetic p-Coumaroyl showing no induction while synthetic C12-HSL did induce. More in-depth figures and data can be found under the Experience tab of this part. | ||
</div> | </div> | ||
Latest revision as of 20:22, 11 October 2018
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Description
A transcription factor, LuxR (BBa_C0062) that is active in the presence of cell-cell signaling molecule 3OC6HSL is controlled by a TetR (BBa_C0040) regulated operator (BBa_R0040). Device input is 3OC6HSL. Device output is PoPS from a LuxR-regulated operator. If used in a cell containing TetR then a second input signal such as [http://openwetware.org/wiki/ATc aTc] can be used to produce a logical AND function. Get the pdf datasheet here.
Performance
| Experiment1 | Characteristic1 | Value1 |
|---|---|---|
| Transfer Function | Maximum Output | 6.6 PoPS cell-1 |
| Hill coefficient | 1.6 | |
| Switch Point | 1.5E-9 M 3OC6HSL, exogenous | |
| Response time: | <1 min | |
| Input compatibility | Strong response to | 3OC6HSL, C6HSL , C7HSL, 3OC8HSL, C8HSL |
| Weak response to | C4HSL, C10HSL, C12HSL | |
| Stability | Genetic Stability (Low/High Input) |
>92/>56 generations |
| Performance Stability (Low/High Input) |
>92/>56 generations | |
| Demand | Internal Demand (Low/High Input) |
Not measured |
| Transcriptional output demand: (Low/High Input) Nt = length of downstream transcript in nucleotides |
(0/6xNt) nucleotides cell-1 s-1 | |
| (0/1.5E-1xNt) RNAP cell-1 | ||
| Growth Rate (Low/High Input) |
54/59 min Doubling time |
1Measured by Ania Labno and Barry Canton 2006-2007
Compatibility
Chassis: Device has been shown to work in BBa_V1000,BBa_V1001,BBa_V1002, E.Coli S30 Extract (data)
Plasmids: Device has been shown to work on pSB3K3 and pSB1A3
Devices: Device has been shown to work with BBa_E0430, BBa_E0434, BBa_E0240
Crosstalk with systems containing BBa_C0040.
Cell-Cell Signaling Systems: Crosstalk with devices using 3OC6HSL, C6HSL, C7HSL, C8HSL, C10HSL.
Contribution: Arizona_State 2016
Authors: Ernesto Luna, Brady Dennison, Cassandra Barrett, Jimmy Xu, Jiaqi Wu, Dr. Karmella Haynes
The 2016 Arizona_State team ran a series of induction tests on 10 AHL Senders (naturally-produced by an BL21 E.coli chassis) and 5 synthetic AHL variants using Bba_F2620 as the output device. This included a series of visual plate induction tests as well as an 8-hour GFP absorbance read. This contribution aims to expand the characterization of this part by identifying crosstalk partners as well as potential orthogonal systems.
The table below summarized the findings of our induction test. Note: these results are not conclusive and are simply what the collected data indicated from our 8-hour induction test.
Two runs at two different concentrations (100nM and 1uM) using synthetic AHLs yielded some interesting differences, with synthetic p-Coumaroyl showing no induction while synthetic C12-HSL did induce. More in-depth figures and data can be found under the Experience tab of this part.