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CapD

Part:BBa_K314011

Designed by: IGEM 2010 Team Washington   Group: iGEM10_Washington   (2010-10-08)
Revision as of 21:51, 26 October 2010 by Wusean (Talk | contribs) (Usage and Biology)

CapD

A protein native to Bacillus anthracis which cleaves poly-D-gamma-glutamate (PDG) and anchors it to the bacterium's peptidoglycan.

Usage and Biology

To test BBa _K314011, we first inserted it into a pET29b+ vector using [http://2010.igem.org/Team:Washington/Protocols/KunkelCapD Kunkel's Mutagenesis]. CapD 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]. A protein gel was also run to determine physical qualities. 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 1540
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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