Difference between revisions of "Part:BBa K4509569"
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
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=Characterization=
 
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==Cell Burden==
 
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J23116 has a burden range of -0.6 ± 11.0%
 
J23116 has a burden range of -0.6 ± 11.0%
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Revision as of 08:01, 12 October 2022


HORSERADISH PEROXIDASE with constitutive promoter J23100

The enzyme horseradish peroxidase, found in the roots of horseradish catalyzes the oxidation of various organic substrates by hydrogen peroxide. It is a metalloenzyme with many isoforms; the most studied type is C. The composite part starts with the BBa_J23100 constitutive promoter, continues with RBS BBa_B0034 and is followed by the Horseradish Peroxidase coding sequence BBa_K1291571. The sequence ends with a terminator and a BamH1 restriction site.

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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 445
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 565
    Illegal AgeI site found at 719
    Illegal AgeI site found at 1022
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

The promoter J23119 is replaced with the promoter J23100 to increase the expression of the enzyme.

hrp-100-1.png

The increased expression of HRP was reviewed by performing the TMB assay.

TMB assay

3,3',5,5'-Tetramethylbenzidine (TMB) is the most commonly used chromogen for horseradish peroxidase (HRP). TMB produces a yellow-orange colour with Horseradish Peroxidase and a stop solution (Sulfuric acid), that absorbs at 450nm.

hrp-expt-1.jpg

Cell Burden

J23116 has a burden range of -0.6 ± 11.0%