Difference between revisions of "Part:BBa K2149020"

 
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===Expression===
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===Biological===
  
We assembled a plasmid with the LuxD gene and a RFP gene to ensure a visual representation of our expression. And the grown bacteria expressed quite a lot of the gene.
 
 
Pelletized cells.Source: Personal archive.
 
So with our visual expression confirmation, we went for the actual confirmation. We then digested our cells and separated our gene from it's plasmid. With our product had both the RFP and the LuxD gene in it's cut product, the relation between the RFP protein and the LuxD protein expressions are then proved.
 
And our gel product proved to be also really intense, with well defined and strong bands. Analyzing the band size, it is also pretty clear the gene composition of it, with around 1740bp for the LuxD gene and 905bp for the RFP, our expression should turn out somewhere between the 2000bp ant 3000bp band, with 2645bp.
 
 
Electrophoresis from expressed LuxD+RFP, with Quick-Load Purple 2-log DNA ladder .Source: Personal archive.
 
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 11:30, 21 October 2016


RFP with Lux D and an aldehyde decarbonylase

The Red Fluorescent Protein (RFP) is a gene reporter used to signalize the expression of Lux D, a transferase which is part of an operon (operon Lux C, E and D) involved in the biosynthesis of alkanes. After the end of gene Lux D there is an aldehyde decarbonylase that catalyzes the conversion of fatty aldehydes, produced by operon Lux, to alkanes.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1897
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1285
    Illegal BamHI site found at 1678
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 555
    Illegal AgeI site found at 667
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1719