Difference between revisions of "Part:BBa K2315011"
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Group: <b>Shanghaitech iGEM 2017</b> | Group: <b>Shanghaitech iGEM 2017</b> | ||
Revision as of 01:34, 31 October 2017
</b>LasR-pLas-GFP</b>
Group: Shanghaitech iGEM 2017
The HSL receiver LasR from P.aeruginosa activates expression of GFP protein in response to 3OC12HSL.
This Receiver can be easily replaced by other AHL receivers in our collection. A full collection could be found in: (link tobe added!!!!!!!!)
Contents
Usage and Biology
When AHL is added with concentration higher than a critical value, the constitutively expressed LasR will bind to the AHL molecule 3OC12HSL, dimerize and bind to the pLas regulatory sequence to activate GFP expression.
Fluorescent Response to cognate 3OC12-HSL
To test this part, we used standard 3OC12HSL (HSL produced by LasI in P.aeruginosa) to determine the response curve.
(Figure2&3-response to 3OC12HSL to be added)
Orthogonality test against non-cognate inducers
We have characterized crosstalk response of LasR to several non-cognate AHLs (Figure4-7-response to non-cognate inducers to be added)
Usages in our Project
We developed a measurement protocol using the fluorescent protein coupled AHL receiver germs to measure the actual AHLs concentration in high precision and sensitivity in compared with LC-MS.
LasR-pLas-GFP
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]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 383
Illegal AgeI site found at 580 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1700