Difference between revisions of "Part:BBa K415000:Design"
 
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<partinfo>BBa_K415000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K415000 SequenceAndFeatures</partinfo>
  
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BBa_K415000 consists of a Plux_cI promoter expressing mCherry. The part was constructed via the insertion of BBa_J06702 into a BBa_R0065 backbone(pSB1A2). The following gel presents confirmation of said construction:
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BBa_K415000 represents an AND-Gate promoter controlling the expression of expressing mCherry. The promoter requires two inputs: an environment free of cI and secondly, quorum sensing, in the form of LuxR-homoserine-lactone for triggering the pLux promoter. The part was constructed via the insertion of BBa_J06702 into a BBa_R0065 backbone(pSB1A2). The following 1% agarose gel, prepared from powder and 1% TAE and stained with SYBR® Safe DNA gel stain, presents confirmation of said construction. The part was run along the standard of Hyperladder 1.
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Figure 1
 
[[Image:K415000 gel.jpg|center|150px]]
 
[[Image:K415000 gel.jpg|center|150px]]
  
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The following is a plasmid map of the part in the pSB1A2 backbone:
 
The following is a plasmid map of the part in the pSB1A2 backbone:
  
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Figure 2
 
[[Image:K415000 geneious.jpg|center|500px]]
 
[[Image:K415000 geneious.jpg|center|500px]]
  
  
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The fluorescence of mCherry was then quantified using FACs(Fluorescence-activated cell sorting) running two samples: -AHL and +AHL. The addition of AHL tests whether or not the circuit will respond without LuxR.  
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The fluorescence of mCherry was then quantified using FACs(Fluorescence-activated cell sorting) running two samples: -AHL and +AHL. The addition of AHL tests whether or not the circuit will respond without LuxR. The fluorescence of mCherry was measured in arbitrary units.
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Figure 3
 
[[Image:K415000.jpg|center|300px]]
 
[[Image:K415000.jpg|center|300px]]
  
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It is clear that the circuit does not respond to the addition of AHL. The following are FACs sheets:
 
It is clear that the circuit does not respond to the addition of AHL. The following are FACs sheets:
  
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-AHL
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Figure 4: -AHL
 
[[Image:K415000Basal.jpg|center|500px]]
 
[[Image:K415000Basal.jpg|center|500px]]
  
  
  
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+AHL
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Figure 5: +AHL
 
[[Image:K415000+AHL facs.jpg|center|500px]]
 
[[Image:K415000+AHL facs.jpg|center|500px]]
  

Latest revision as of 03:57, 26 October 2010

pLux/cI-OR : RBS-mCherry : Term


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

BBa_K415000 represents an AND-Gate promoter controlling the expression of expressing mCherry. The promoter requires two inputs: an environment free of cI and secondly, quorum sensing, in the form of LuxR-homoserine-lactone for triggering the pLux promoter. The part was constructed via the insertion of BBa_J06702 into a BBa_R0065 backbone(pSB1A2). The following 1% agarose gel, prepared from powder and 1% TAE and stained with SYBR® Safe DNA gel stain, presents confirmation of said construction. The part was run along the standard of Hyperladder 1.

Figure 1

K415000 gel.jpg


The following is a plasmid map of the part in the pSB1A2 backbone:

Figure 2

K415000 geneious.jpg


The fluorescence of mCherry was then quantified using FACs(Fluorescence-activated cell sorting) running two samples: -AHL and +AHL. The addition of AHL tests whether or not the circuit will respond without LuxR. The fluorescence of mCherry was measured in arbitrary units.

Figure 3

K415000.jpg


It is clear that the circuit does not respond to the addition of AHL. The following are FACs sheets:

Figure 4: -AHL

K415000Basal.jpg


Figure 5: +AHL

K415000+AHL facs.jpg



Source

https://parts.igem.org/wiki/index.php?title=Part:BBa_R0065

https://parts.igem.org/wiki/index.php?title=Part:BBa_J06702

References