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CapH

Part:BBa_K314015

Designed by: Chris Eiben, Tomas Huber, David Zong   Group: iGEM10_Washington   (2010-10-08)
Revision as of 20:43, 27 October 2010 by Wusean (Talk | contribs) (Usage and Biology)

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CapD_CP-F24H

A mutated [http://www.parts.igem.org/Part:BBa_K314012 CapD_CP] protein aimed at increasing hydrolysis and decreasing transpeptidation of PDG.

Usage and Biology

To test BBa _K314015, it was inserted into a pET29b+ vector using [http://2010.igem.org/Team:Washington/Protocols/KunkelCapD Kunkel Mutagenesis]. CapD_CP-F24H was then produced and purified as described in the [http://2010.igem.org/Team:Washington/Protocols/50mLPurificationCapD UW 2010 iGEM Team's Small Scale Protein Purification Protocol]. The purified protein was then tested for activity against CapD_CP. For a detailed description of the assay, please see the [http://2010.igem.org/Team:Washington/Protocols/EnzymeAssayCapD 2010 UW iGEM wiki]. The resulting data is shown below.

Kinetic constants, kcat and Km, taken from our plotted curves are shown in the table above.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1489
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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