Difference between revisions of "Part:BBa K1455002"
 
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<partinfo>BBa_K1455002 short</partinfo>
 
<partinfo>BBa_K1455002 short</partinfo>
  
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This device is used to express LuxO D47E by arabinose induction and  and at the same time to measure the expression of LuxOD 47E  in an indirectly way by GFP fluoresence.  
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This device is used to express LuxO D47E by arabinose induction and at the same time to measure the expression of LuxOD 47E  in an indirectly way by GFP fluoresence.  
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LuxOD 47E acts like a phosphorilated LuxO, which is an essential protein of ''Vibrio spp.''  quorum sensing system. LuxO is the final protein in a phophorilation cascade and binds, as a trancription facotr, to pqrr promoters. LuxOD 47E is able to activate the genes downstream of a pqrr promoter.
 
LuxOD 47E acts like a phosphorilated LuxO, which is an essential protein of ''Vibrio spp.''  quorum sensing system. LuxO is the final protein in a phophorilation cascade and binds, as a trancription facotr, to pqrr promoters. LuxOD 47E is able to activate the genes downstream of a pqrr promoter.
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Arabinose inducible promoter used here is a strong one.
  
 
The following pictures were taken using a Eclipse-Ti Nikon® ECLIPSE Ti inverted microscope equiped with an ANDOR iXon Ultra 897 camera. The filter used was FITC.
 
The following pictures were taken using a Eclipse-Ti Nikon® ECLIPSE Ti inverted microscope equiped with an ANDOR iXon Ultra 897 camera. The filter used was FITC.
  
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Here is ''E. coli'' DH5α carrying this device after one hour of induction with 0.1 % arabinose: light (left) and FITC (right)
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Here is ''E. coli'' DH5α carrying this device after one hour of induction with 0.1 % arabinose, light (left) and FITC (right):
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[[File:unahoraraluz.png|left|300px]]
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[[File:unahoraarafitc.png|right|300px]]
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[[File:unahoraraluz.png|350px]]
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[[File:unahoraarafitc.png|350px]]
  
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And after 3 hours:
 
  
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[[File:treshorasara.png|300px]]
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And after 3 hours (both with FITC):
  
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[[File:treshorasara.png|350px]]
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[[File:treshorasara2.png|350px]]
  
  

Latest revision as of 02:18, 8 October 2014

pBAD - luxO D47E - GFP

This device is used to express LuxO D47E by arabinose induction and at the same time to measure the expression of LuxOD 47E in an indirectly way by GFP fluoresence.

LuxOD 47E acts like a phosphorilated LuxO, which is an essential protein of Vibrio spp. quorum sensing system. LuxO is the final protein in a phophorilation cascade and binds, as a trancription facotr, to pqrr promoters. LuxOD 47E is able to activate the genes downstream of a pqrr promoter.

Arabinose inducible promoter used here is a strong one.

The following pictures were taken using a Eclipse-Ti Nikon® ECLIPSE Ti inverted microscope equiped with an ANDOR iXon Ultra 897 camera. The filter used was FITC.

Here is E. coli DH5α carrying this device after one hour of induction with 0.1 % arabinose, light (left) and FITC (right):

Unahoraraluz.png Unahoraarafitc.png


And after 3 hours (both with FITC):

Treshorasara.png Treshorasara2.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
    Illegal XhoI site found at 292
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 980
    Illegal BsaI.rc site found at 2189
    Illegal SapI.rc site found at 942