Difference between revisions of "Team:ETH Zurich/Parts"
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<img src="https://static.igem.org/mediawiki/2015/thumb/0/04/ETH_favpart.png/120px-ETH_favpart.png" style="position:absolute;left:-6%;top:-40%;pointer-events: none;"> | <img src="https://static.igem.org/mediawiki/2015/thumb/0/04/ETH_favpart.png/120px-ETH_favpart.png" style="position:absolute;left:-6%;top:-40%;pointer-events: none;"> | ||
<p><a href="http://parts.igem.org/Part:BBa_K1847010">BBa_K1847010</a>. Hybrid promoter that is responsive to both IPTG and lactate.</p> | <p><a href="http://parts.igem.org/Part:BBa_K1847010">BBa_K1847010</a>. Hybrid promoter that is responsive to both IPTG and lactate.</p> | ||
| − | <p><a href="http://parts.igem.org/Part:BBa_K1847008">BBa_K2847008</a>. Promoter that shows an improved performance when compared | + | <p><a href="http://parts.igem.org/Part:BBa_K1847008">BBa_K2847008</a>. Promoter that shows an <b>improved performance</b> when compared to the natural <a href="http://parts.igem.org/Part:BBa_K822000">P<sub>lldR</sub></a></p> |
</div> | </div> | ||
<p></p> | <p></p> | ||
Revision as of 03:08, 19 September 2015
- Project
- Modeling
- Lab
- Human
Practices - Parts
- About Us
Parts
Many parts were required to complete our project. Some of these parts where used from the Registry, others were designed and cloned for this project. In the following section, we are presenting all parts we used, designed and characterized.
Overview of our genetic design.
BBa_K1847010. Hybrid promoter that is responsive to both IPTG and lactate.
BBa_K2847008. Promoter that shows an improved performance when compared to the natural PlldR
Our part collection
We designed a promoter library based on the wild-type PlldR (BBa_K822000). You can check our results using this promoters here.
Regulatory system design |
Content |
BioBrick |
|---|---|---|
 |
Promoter designed keeping the original architecture and changing the wild-type promoter by a strong Anderson promoter (BBa_J23100) | BBa_K1847007 |
 |
Promoter designed keeping the original architecture and changing the wild-type promoter by a weak Anderson promoter (BBa_J23117) | BBa_K1847008 |
 |
Promoter designed keeping the original architecture and changing the wild-type promoter by a medium Anderson promoter (BBa_J23118) | BBa_K1847009 |
 |
Promoter designed substituting the promoter by a non-functional DNA sequence, with an Anderson promoter placed in front of the operators (BBa_J23117) | BBa_K1847005 |
 |
Promoter designed substituting the promoter by a non-functional DNA sequence, with an Anderson promoter placed in front of the operators (BBa_J23118) | BBa_K1847006 |
 |
In this design, the spacing between the two operator sites was removed, and the Anderson promoter was placed in front of the operators (BBa_J23100) | BBa_K1847002 |
 |
In this design, the spacing between the two operator sites was removed, and the Anderson promoter was placed in front of the operators (BBa_J23117) | BBa_K1847003 |
 |
In this design, the spacing between the two operator sites was removed, and the Anderson promoter was placed in front of the operators (BBa_J23118) | BBa_K1847004 |
 |
The original PlldR was substituted by Plac and lacO. | BBa_K1847010 |
 |
The original PlldR was substituted by PlacUV5 and lacO. | BBa_K1847011 |
 |
The original PlldR was substituted by a spacer and Plac and lacO were located in front of the promoter. | BBa_K1847012 |
 |
Plac and lacO | BBa_K1847013 |
 |
PlacUV5 and lacO | BBa_K1847014 |
Our new basic parts
In this section we present LldR and LldP, the regulator protein and transporter protein of the LldPRD operon, respectively, as well as a synthetically designed hybrid promoter responsive to lactate and IPTG.
Part |
Content |
Registry number |
|---|---|---|
| LldR | Sequence encoding the regulator protein LldR. | BBa_K1847001 |
| LldP-LldR | Sequence encoding the L-lactate permease LldP and the regulator protein LldR. | BBa_K1847016 |
| Synthetically designed hybrid promoter | Synthetic promoter inhibited by lactate and IPTG. | BBa_K1847010 |
Our new composite parts
Our composite parts are from used in binding system to E. coli. More information about our experiments on bacterial binding to mammalian cells can be found here.
Part |
Content |
Registry number |
|---|---|---|
| Annexin V | Promoter J23114-RBS B0034- Annexin V optimized for Escherichia coli | BBa_K1847000 |
| INP-Annexin V | Promoter J23114-RBS B0034- INP-Annexin V fusion protein | BBa_K1847015 |
Newly characterized parts of the Registry
Here we present the parts from the Registry where we improved the existing characterization.
Part Name |
Description |
Registry Number |
|---|---|---|
| aiiA | Autoinducer inactivation enzyme aiiA (enzyme that inactivates the acylhomoserine lactone (AHL) quorum-sensing signal). Check out our characterization for aiiA! | BBa_C0160 |
| pLldR | lldPRD operon promoter + RBS from E. coli. In the absence of L-lactate, lldR binds to two operator sites O1 and O2 in the promoter region and inhibits expression. Check out our characterization for pLldR! We also improved the existing promoter by removing the ArcA site and obtained an increased ON/OFF ratio by changing the promoter strength. | BBa_K822000 |
Used parts from of the Registry
In this section we present other parts from the Registry that we used during our project.
Part Name |
Description |
Registry Number |
INP-EYFP | Fusion of Ice Nucleation Protein (INP) and Enhanced Yellow Fluorescent Protein (EYFP). Check out our use of INP! | BBa_K523013 |
|---|---|---|
| lacI + LVA | lacI repressor from E. coli (+LVA) | BBa_C0012 |
| pLuxR | Promoter activated by the LuxR protein complexed with the autoinducer homoserine lactone (HSL) | BBa_R0062 |
| cI | cI repressor from E. coli phage lambda modified with an LVA tail for rapid degradation of the protein | BBa_C0051 |
| HylA | HlyA-tag+Secretion system, allows a protein to be secreted by means of the alpha-hemolysin secretion system in E. coli | BBa_K1166002 |
| InterLab 1 strong promoter | Anderson promoter J23101 from the Anderson collection | BBa_K823005 (BBa_J23101) |
| InterLab 2 medium-strong promoter | Anderson promoter J23106 from the Anderson collection | BBa_K823008 (BBa_J23106) |
| terminator | Transcription terminator for the E.coli RNA polymerase | BBa_B0012 |
| double terminator | Double terminator consisting of BBa_B0010 and BBa_B0012 | BBa_B0015 |
| very strong promoter | Strong member of the family of promoters J23100 through J23119 | BBa_J23100 |
| gfp (InterLab) | intermediate in screening plasmid construction containing GFP | BBa_I13504 |
| medium promoter | Medium member of the family of promoters J23100 through J23119 | BBa_J23118 |
| LuxI | autoinducer synthetase for acylhomoserine lactone (AHL), no LVA | BBa_C0161 |
| InterLab 3 weak promoter | Anderson promoter J23117 from the Anderson collection | BBa_K823013 (BBa_J23117) |
| promoter medium-weak | Medium-weak member of the family of promoters J23100 through J23119 | BBa_J23114 |
| RBS | RBS.3 (medium) derivative of BBa_0030 | BBa_B0032 |
| promoter plus GFP (InterLab control) | J23151 inserted in the Promoter MeasKit | BBa_I20270 |
| terminator | Artificial terminator, estimated %T~>90% | BBa_B1006 |
We would like to thank our sponsors
