Difference between revisions of "Part:BBa K415000"

 
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K415000 short</partinfo>
 
<partinfo>BBa_K415000 short</partinfo>
  
−
This is a BioBrick composed of parts BBa_R0065 and BBa_J06702. This composite produces the mCherry fluorescent protein. It is activated by the presence of LuxR and AHL, and it is repressed by cI.
+
<partinfo>BBa_K415000 SequenceAndFeatures</partinfo>
  
−
<!-- Add more about the biology of this part here
+
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.
−
===Usage and Biology===
+
  
−
<!-- -->
+
Figure 1
−
<span class='h3bb'>Sequence and Features</span>
+
[[Image:K415000 gel.jpg|center|150px]]
−
<partinfo>BBa_K415000 SequenceAndFeatures</partinfo>
+
 
 +
 
 +
The following is a plasmid map of the part in the pSB1A2 backbone:
 +
 
 +
Figure 2
 +
[[Image:K415000 geneious.jpg|center|500px]]
 +
 
 +
 
 +
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
 +
[[Image:K415000.jpg|center|300px]]
 +
 
 +
 
 +
It is clear that the circuit does not respond to the addition of AHL. The following are FACs sheets:
 +
 
 +
Figure 4: -AHL
 +
[[Image:K415000Basal.jpg|center|500px]]
 +
 
 +
 
 +
 
 +
Figure 5: +AHL
 +
[[Image:K415000+AHL facs.jpg|center|500px]]
 +
 
 +
 
 +
 
 +
 
 +
===Source===
 +
 
 +
https://parts.igem.org/wiki/index.php?title=Part:BBa_R0065
  
 +
https://parts.igem.org/wiki/index.php?title=Part:BBa_J06702
  
−
<!-- Uncomment this to enable Functional Parameter display
+
===References===
−
===Functional Parameters===
+
−
<partinfo>BBa_K415000 parameters</partinfo>
+
−
<!-- -->
+

Revision as of 04:04, 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