Team:British Columbia/pBAD

From 2009.igem.org

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(Overview: Added data, colored sequences)
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In order to make an analog biosensor, we need our traffic light to be able to respond differently to different concentrations of an input. However, the Registry is lacking in variable strength inducible promoters. We designed two variants of the <partinfo>I13453</partinfo> pBAD promoter, one weaker and one stronger, based on AraC binding experiments performed by [[Team:British_Columbia/Bibliography|Niland et al.]]
In order to make an analog biosensor, we need our traffic light to be able to respond differently to different concentrations of an input. However, the Registry is lacking in variable strength inducible promoters. We designed two variants of the <partinfo>I13453</partinfo> pBAD promoter, one weaker and one stronger, based on AraC binding experiments performed by [[Team:British_Columbia/Bibliography|Niland et al.]]
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  pbad wt      ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGC'''ATTTTTA'''TCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
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  pbad wt      ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGC<font color=red>ATTTTTA</font>TCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
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  pbad weak    ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGC'''TTTTTTA'''TCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
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  pbad weak    ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGC<font color=red>TTTTTTA</font>TCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
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  pbad strong  ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGC'''AAGATAG'''TCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
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  pbad strong  ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGC<font color=red>AAGATAG</font>TCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
Above is a sequence alignment of our promoter variants with the wild type pBAD, truncated for space. For the full sequences, see our parts in the registry at <partinfo>K206000</partinfo> (strong) and <partinfo>K206001</partinfo> (weak).
Above is a sequence alignment of our promoter variants with the wild type pBAD, truncated for space. For the full sequences, see our parts in the registry at <partinfo>K206000</partinfo> (strong) and <partinfo>K206001</partinfo> (weak).
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<html>Here is the experimental data regarding the strength of these promoters when coupled to RFP: <a href="https://static.igem.org/mediawiki/2009/e/e6/British_Columbia_pBAD_RFP%2BTimecourse%2B3.xls">pBAD_RFP_Timecourse_3.xls</a></html>

Revision as of 07:37, 20 October 2009

IGEMFinalGoldDNA-logo.jpg Home The Team The Traffic Light pBAD Promoters Lock and Key Jammer Parts Submitted to the Registry Our Sponsors Lab Notebook Bibliography

Overview

In order to make an analog biosensor, we need our traffic light to be able to respond differently to different concentrations of an input. However, the Registry is lacking in variable strength inducible promoters. We designed two variants of the pBAD promoter, one weaker and one stronger, based on AraC binding experiments performed by Niland et al.

pbad wt      ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCATTTTTATCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
pbad weak    ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCTTTTTTATCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
pbad strong  ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGATAGTCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...

Above is a sequence alignment of our promoter variants with the wild type pBAD, truncated for space. For the full sequences, see our parts in the registry at (strong) and (weak).

Here is the experimental data regarding the strength of these promoters when coupled to RFP: pBAD_RFP_Timecourse_3.xls