Difference between revisions of "Part:BBa K1615067"

 
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Laccase TVEL5 from Tramates versicolor.  
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Laccases are glycosylated polyphenol oxidases and have four copper ions per molecule<sup>1</sup>. This allows them to catalyse the reduction of O<sub>2</sub> to 2H<sub>2</sub>O while oxidising an aromatic substrate<sup>2</sup>. This laccase is coded by the structural gene <i>lcc</i> in <i>Trametes versicolor</i>, a species of white rot fungus<sup>3</sup>.
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To increase the functions of laccase, we took the sequence from iGEM12_Bielefeld-Germany laccase BBa_K863030 and codon optimised it for the chassis <i>Escherichia coli</i>. We then made it RFC25 compatible by adding the prefix and suffix and removing all illegal restriction sites. This laccase can now be used in protein fusions as there is an in-frame 6-nucleotide scar.
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<sup>1</sup>Lontie, R. (1984). <i>Copper proteins and copper enzymes</i> (Vol. 2). CRC.
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<sup>2</sup>Thurston, C. F. (1994). The structure and function of fungal laccases. <i>Microbiology</i>, 140(1), 19-26.
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<sup>3</sup>Collins, P. J., & Dobson, A. (1997). Regulation of laccase gene transcription in Trametes versicolor. <i>Applied and Environmental Microbiology</i>, 63(9), 3444-3450.
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Revision as of 23:04, 21 September 2015

Laccase TVEL5 in RFC25

Laccases are glycosylated polyphenol oxidases and have four copper ions per molecule1. This allows them to catalyse the reduction of O2 to 2H2O while oxidising an aromatic substrate2. This laccase is coded by the structural gene lcc in Trametes versicolor, a species of white rot fungus3.

To increase the functions of laccase, we took the sequence from iGEM12_Bielefeld-Germany laccase BBa_K863030 and codon optimised it for the chassis Escherichia coli. We then made it RFC25 compatible by adding the prefix and suffix and removing all illegal restriction sites. This laccase can now be used in protein fusions as there is an in-frame 6-nucleotide scar.


1Lontie, R. (1984). Copper proteins and copper enzymes (Vol. 2). CRC. 2Thurston, C. F. (1994). The structure and function of fungal laccases. Microbiology, 140(1), 19-26. 3Collins, P. J., & Dobson, A. (1997). Regulation of laccase gene transcription in Trametes versicolor. Applied and Environmental Microbiology, 63(9), 3444-3450.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 612
    Illegal BglII site found at 773
    Illegal BamHI site found at 836
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 418