Difference between revisions of "Part:BBa K1073030"
(→Induction of expression by RhlR and N-3-oxododecanoyl-HSL) |
|||
| Line 10: | Line 10: | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-buturyl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-buturyl-HSL to the culture broth. | + | The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-buturyl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-buturyl-HSL to the culture broth. Expression of ''lasR'' is regulated by the strong constitutive promoter <bbpart>BBa_J23100</bbpart> and RBS of medium strengh (<bbpart>BBa_B0032</bbpart>). |
Note that due to the leakiness of the promoter there is background activity of the beta lactamase even when N-3-oxododecanoyl-HSL is not present. | Note that due to the leakiness of the promoter there is background activity of the beta lactamase even when N-3-oxododecanoyl-HSL is not present. | ||
Revision as of 18:07, 24 September 2013
Inducible ampicillin resistance cassette + LasR expression cassette
Induction of the beta lactamase mediated ampicillin resistance by LasR and specific autoinducer N-3-oxododecanoyl homoserine lactone.
In complex with the autoinducer molecule N-3-oxododecanoyl homoserine lactone (N-3-oxododecanoyl-HSL) the transcription regulator LasR binds to specific sequences in the promoter region and enables transcription of ampR. This construct provides all the parts required to express the gene downstream of the inducible promoter. Only LasI, the synthase producing the autoinducer molecule N-3-oxododecanoyl-HSL is missing in the construct. However, N-3-oxododecanoyl-HSL can be taken up from the media.
Usage and Biology
The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-buturyl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-buturyl-HSL to the culture broth. Expression of lasR is regulated by the strong constitutive promoter BBa_J23100 and RBS of medium strengh (BBa_B0032).
Note that due to the leakiness of the promoter there is background activity of the beta lactamase even when N-3-oxododecanoyl-HSL is not present.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1169
Illegal NheI site found at 1192 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1546
Illegal AgeI site found at 1743 - 1000COMPATIBLE WITH RFC[1000]