Part:BBa_K2315011

Rpa-Las signal converter

Group: Shanghaitech iGEM 2017

Introduction

HSL receiver from P.aeruginosa, actives expression of GFP protein in response to 3OC12HSL.

(Figure1-mechanism to be added)

When proper AHL added with concentration higher than a critical value, The constitutively expressed LasR will bind the signaling AHL molecule, dimerize and bind to pLas regulatory sequence before GFP gene and actives its expression.

This part is one of the interchangeable AHL receivers. Full collection could be viewed at:

Fluorescent Response cognate 3OC12HSL produced by LasI in P.aeruginosa

To validate the part's design, we've used standard 3OC12HSL (HSL produced by LasI in P.aeruginosa) sample to determine the response curve of this part.

(Figure2&3-response to 3OC12HSL to be added)

Orthogonality test against non-cognate inducers

(Figure4-7-response to non-cognate inducers to be added)

Usage: Measuring unknown AHL production

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

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 383
  Illegal AgeI site found at 580
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1700