Team:Chiba/AHL Receiver Phase

From 2008.igem.org

(Difference between revisions)
(Receiver)
(Receiver)
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*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002 (Express GFP in response to AHL)]
*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002 (Express GFP in response to AHL)]
[[Image:High-Copy-Receiver Chiba.gif]]<br clear=all>
[[Image:High-Copy-Receiver Chiba.gif]]<br clear=all>
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*Low Copy Receiver
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[[Image:Low-Copy-Receiver Chiba.gif]]
===Result===
===Result===

Revision as of 16:26, 28 October 2008

Chiba-U.gif

Home The Team The Project Parts Submitted to the Registry Reference Notebook Acknowledgements

Signal Molecule Receiver Phase

Quorum Sensing Crosstalk

Design

Table.1 LuxR family gene

センダーとレシーバーを元来とは異なる組み合わせにすることによって、thresholdに達する時間を遅くする。(杉山)


Experiment

  • Sender
    • AHL autoinucer[http://partsregistry.org/Part:BBa_S03623 (BBa_S03623) ]
LuxI-sender Chiba.gif

Fig.R family crosstalk test
  • Receivers
    • LuxR

LuxR-p15A Chiba.gif

    • RhlR[http://partsregistry.org/Part:BBa_K084004 (BBa_K084004)]

RhlR-p15A Chiba.gif

    • LasR[http://partsregistry.org/Part:BBa_K084005 (BBa_K084005)]

LasR-p15A Chiba.gif

    • CinR[http://partsregistry.org/Part:BBa_K084006 (BBa_K084006)]

CinR-p15A Chiba.gif

  • Reporter
    • Plux-GFP[http://partsregistry.org/Part:BBa_J37032 (BBa_J37032)]

Plux-GFP-pMB1 Chiba.gif



Result

Fig.  Time Delay Test




Plasmid Copy number

Design

Experiment

Sender

  • AHL autoinucer[http://partsregistry.org/Part:BBa_S03623 (BBa_S03623) ]
LuxI-sender Chiba.gif

Receiver

  • [http://partsregistry.org/Part:BBa_T9002 BBa_T9002 (Express GFP in response to AHL)]

High-Copy-Receiver Chiba.gif

  • Low Copy Receiver

Low-Copy-Receiver Chiba.gif

Result

Fig.  Time Delay Test


LuxR_mutant

Design

Fig.6 Location of the LuxR mutant in LuxR
Table.1 Seisitivity of LuxR mutants. Data modified from Collins et.al. Mol. Microbiol. 55, 712–723 (2005)


Collins et.al. described the hyper-sensitive variants of luxR to AHL.(Collins, C. H., Arnold, F. H. & Leadbetter, J. R. Directed evolution of Vibrio fischeri LuxR for increased sensitivity to a broad spectrum of acyl-homoserine lactones. Mol. Microbiol. 55, 712–723 (2005))

私たちはmutated Receiverを用いることで、AHL感受性の違う2種類(WTと変異体)のレシーバーを用意し、delay-timeのバリエーションを増やした。 (小林)


Experiment

Result

Home The Team The Project Parts Submitted to the Registry Reference Notebook Acknowledgements