Difference between revisions of "Part:BBa K2315011"

Revision as of 14:18, 30 October 2017

Introduction

(Figure1-mechanism to be added)

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)

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

@@ Line 2: / Line 2: @@
 Group: <b>Shanghaitech iGEM 2017</b>
-HSL receiver from P.aeruginosa, actives expression of GFP protein in response to 3OC12HSL.
+The HSL receiver LasR from P.aeruginosa activates expression of GFP protein in response to 3OC12HSL.
-This part is one of the interchangeable AHL receivers. Full collection could be viewed at:
+This Receiver can be easily replaced by other AHL receivers in our collection. A full collection could be found in: (link tobe added!!!!!!!!)
 ==<b>Introduction</b>==
@@ Line 10: / Line 10: @@
 (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.
+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.
 ==<b>Fluorescent Response to cognate 3OC12-HSL</b>==
-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.
+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've characterized the crosstalk between LasR and non-cognate AHLs
+We have characterized crosstalk response of LasR to several non-cognate AHLs
 (Figure4-7-response to non-cognate inducers to be added)