Difference between revisions of "Part:BBa K2267044"
(→‎result)
 
(2 intermediate revisions by the same user not shown)
Line 10: Line 10:
 
<!-- -->
 
<!-- -->
 
Quorum sensing is a naturally occurring mechanism that certain strains of bacteria use to regulate gene expression in response to their population density. These bacteria secrete autoinducer signalling molecules, such as N-acyl homoserine lactones (AHLs), that bind to transcription factors to alter gene expression.
 
Quorum sensing is a naturally occurring mechanism that certain strains of bacteria use to regulate gene expression in response to their population density. These bacteria secrete autoinducer signalling molecules, such as N-acyl homoserine lactones (AHLs), that bind to transcription factors to alter gene expression.
−
In this case, the constitutively expressed LuxR transcriptional regulator (C0062) is activated by the binding of 3O-C6 AHL, an AHL quorum signal. The activated LuxR regulator binds to a LuxR-inducible promoter, pLux, upstream of a GFP reporter. As a result, GFP is expressed when the receiver device is induced with 3O-C6 AHL. We designed this part to characterize the activation range of the Lux receiver device.
+
In this case, the constitutively expressed LuxR transcriptional regulator ( https://parts.igem.org/Part:BBa_K2267032 we improved) is activated by the binding of 3O-C6 AHL, an AHL quorum signal. The activated LuxR regulator binds to a LuxR-inducible promoter, pLux, upstream of a GFP reporter. As a result, GFP is expressed when the receiver device is induced with 3O-C6 AHL. We designed this part to characterize the activation range of the Lux receiver device.
  
−
  https://parts.igem.org/Part:BBa_K2267026 is Control，  
+
  https://parts.igem.org/Part:BBa_K2267040 is Control，  
−
 
+
−
part1: https://parts.igem.org/Part:BBa_K2267026+https://parts.igem.org/Part:BBa_K2267029+https://parts.igem.org/Part:BBa_K319039.
+
−
 
+
−
part3: https://parts.igem.org/Part:BBa_K2267026+ https://parts.igem.org/Part:BBa_K2267030+https://parts.igem.org/Part:BBa_K319039.
+
−
 
+
−
part4: https://parts.igem.org/Part:BBa_K2267026+ https://parts.igem.org/Part:BBa_K2267031+https://parts.igem.org/Part:BBa_K319039.
+
−
 
+
−
part6: https://parts.igem.org/Part:BBa_K2267026+ https://parts.igem.org/Part:BBa_K2267032+https://parts.igem.org/Part:BBa_K319039.
+
−
 
+
−
part8: https://parts.igem.org/Part:BBa_K2267026+ https://parts.igem.org/Part:BBa_K2267033+https://parts.igem.org/Part:BBa_K319039.
+
  
 
part1:  https://parts.igem.org/Part:BBa_K2267043  
 
part1:  https://parts.igem.org/Part:BBa_K2267043  
Line 35: Line 25:
  
 
===result===
 
===result===
 +
we did point mutation on https://parts.igem.org/Part:BBa_R0062 and the exact spots are
 +
 +
plux1: https://parts.igem.org/Part:BBa_K2267029
 +
 +
plux3: https://parts.igem.org/Part:BBa_K2267030
 +
 +
plux4: https://parts.igem.org/Part:BBa_K2267031
 +
 +
plux6: https://parts.igem.org/Part:BBa_K2267032
 +
 +
plux8: https://parts.igem.org/Part:BBa_K2267046
 +
 +
listed below.
 +
We linked gfp reporter genes
 +
luxr-plux1-gfp: https://parts.igem.org/Part:BBa_K2267043
 +
 +
luxr-plux3-gfp: https://parts.igem.org/Part:BBa_K2267041
 +
 +
luxr-plux4-gfp: https://parts.igem.org/Part:BBa_K2267042
 +
 +
luxr-plux6-gfp: https://parts.igem.org/Part:BBa_K2267044
 +
 +
luxr-plux8-gfp: https://parts.igem.org/Part:BBa_K2267045
 +
 +
on them respectively to test the function of modified promoters.
  
 
https://static.igem.org/mediawiki/2017/e/e6/T--TUST_China--part_part_results.png
 
https://static.igem.org/mediawiki/2017/e/e6/T--TUST_China--part_part_results.png

Latest revision as of 03:38, 2 November 2017


LuxR-I-Plux-GFP-6

This composite part is based on the Lux quorum-sensing system from Vibrio fischerii. It consists of a Lux receiver device and a GFP reporter that is activated in the presence of a 3O-C6 AHL signal.

Quorum sensing is a naturally occurring mechanism that certain strains of bacteria use to regulate gene expression in response to their population density. These bacteria secrete autoinducer signalling molecules, such as N-acyl homoserine lactones (AHLs), that bind to transcription factors to alter gene expression. In this case, the constitutively expressed LuxR transcriptional regulator ( https://parts.igem.org/Part:BBa_K2267032 we improved) is activated by the binding of 3O-C6 AHL, an AHL quorum signal. The activated LuxR regulator binds to a LuxR-inducible promoter, pLux, upstream of a GFP reporter. As a result, GFP is expressed when the receiver device is induced with 3O-C6 AHL. We designed this part to characterize the activation range of the Lux receiver device. 

https://parts.igem.org/Part:BBa_K2267040  is Control，

part1: https://parts.igem.org/Part:BBa_K2267043

part3: https://parts.igem.org/Part:BBa_K2267041

part4: https://parts.igem.org/Part:BBa_K2267042

part6: https://parts.igem.org/Part:BBa_K2267044

part8: https://parts.igem.org/Part:BBa_K2267045

result

we did point mutation on https://parts.igem.org/Part:BBa_R0062 and the exact spots are

plux1: https://parts.igem.org/Part:BBa_K2267029

plux3: https://parts.igem.org/Part:BBa_K2267030

plux4: https://parts.igem.org/Part:BBa_K2267031

plux6: https://parts.igem.org/Part:BBa_K2267032

plux8: https://parts.igem.org/Part:BBa_K2267046

listed below. We linked gfp reporter genes luxr-plux1-gfp: https://parts.igem.org/Part:BBa_K2267043

luxr-plux3-gfp: https://parts.igem.org/Part:BBa_K2267041

luxr-plux4-gfp: https://parts.igem.org/Part:BBa_K2267042

luxr-plux6-gfp: https://parts.igem.org/Part:BBa_K2267044

luxr-plux8-gfp: https://parts.igem.org/Part:BBa_K2267045

on them respectively to test the function of modified promoters.

T--TUST_China--part_part_results.png 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
    Illegal NheI site found at 837
    Illegal NheI site found at 860
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 928
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1452
    Illegal BsaI.rc site found at 2179