Difference between revisions of "Part:BBa K2116025"
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− | An esabox is an 18bp sequence to which the transcriptional regulator EsaR can bind. We placed one esabox right after the normally constitutive Anderson promoter [[Part: | + | An esabox is an 18bp sequence to which the transcriptional regulator EsaR can bind. We placed one esabox right after the normally constitutive Anderson promoter [[Part:BBa_J23118]] to create an EsaR repressible promoter. Transcription can be initiated by the specific AHL EsaR responds to [N-(3-oxo-hexanoyl)-L-homoserine lactone]. |
When characterising this part, we used the D91G variant of EsaR, which has been documented to be more responsive to lower AHL concentrations compared to the wild type EsaR [1]. This EsaR variant was obtained through addgene, and can be found on the registry [[Part:BBa_K2116001]]. | When characterising this part, we used the D91G variant of EsaR, which has been documented to be more responsive to lower AHL concentrations compared to the wild type EsaR [1]. This EsaR variant was obtained through addgene, and can be found on the registry [[Part:BBa_K2116001]]. |
Revision as of 22:40, 16 October 2016
EsaR repressible promoter
An esabox is an 18bp sequence to which the transcriptional regulator EsaR can bind. We placed one esabox right after the normally constitutive Anderson promoter Part:BBa_J23118 to create an EsaR repressible promoter. Transcription can be initiated by the specific AHL EsaR responds to [N-(3-oxo-hexanoyl)-L-homoserine lactone].
When characterising this part, we used the D91G variant of EsaR, which has been documented to be more responsive to lower AHL concentrations compared to the wild type EsaR [1]. This EsaR variant was obtained through addgene, and can be found on the registry Part:BBa_K2116001.
Two copies of EsaR were placed under the control of individual constitutive promoters [Bba_J23118] and expressed on a medium-low copy plasmid (replication origin pBR322/rop). Based on the 2013 paper by Shong&Collins we decided to test AHL concentrations in the range of 0 to 10 000nM, since in all combinations of esabox placements tested in this paper showed saturation at 10 000nM as seen below:
We observed EsaR response to the same AHL, but did not see the same saturation behaviour.
References:
[1] Shong, Jasmine and Cynthia H. Collins. "Engineering The Esar Promoter For Tunable Quorum Sensing-Dependent Gene Expression". ACS Synth. Biol. 2.10 (2013): 568-575.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]