Difference between revisions of "Part:BBa K404205"

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The BAP (Biotinylation Acceptor Peptide) that we included in our Virus Construction Kit is a 15 amino acid long peptide identified by Schatz J., 1993 in an library screening approach and published under the number #85. This peptide with the sequence 5' - GLNDIFEAQKIEWHE - 3' contains a central lysine that is specifically biotinylated by the prokaryotic enzyme biotin holenzyme synthetase, encoded in the BirA gene of E. coli. Specific biotinylation of this peptide sequence can be performed in vivo by contransfecting a plasmid with the BirA gene as described for the AAV in Arnold et al.; 2006 or by an in vitro coupling approach using the purified Escherichia coli enzyme biotin ligase (BirA). <br>
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The purified BirA biotin ligase that was kindly provided from Avidity.com
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The Biotinylation Acceptor Peptide (BAP) is a 15 amino acid long peptide identified by Schatz J., 1993 in an library screening approach. This peptide with the sequence 5' - GLNDIFEAQKIEWHE - 3' contains a central lysine that is specifically biotinylated by the prokaryotic enzyme biotin holenzyme synthetase, encoded in the BirA gene of E. coli. Specific biotinylation of this peptide sequence can be performed in vivo by contransfecting a plasmid with the BirA gene as described for the AAV in Arnold et al.; 2006 or by an in vitro coupling approach using the purified Escherichia coli enzyme biotin ligase (BirA).  
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<h3>References:</h3>
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<div style="float:right; width:480px; height:auto; "><img src="https://static.igem.org/mediawiki/parts/a/a9/Freiburg10_ViralBrick_motif_BAP.png" width="460"  
 
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Revision as of 17:47, 26 October 2010

ViralBrick-587-BAP
The Biotinylation Acceptor peptide motif, ready for insertion into the 587 loop

Freiburg10 ViralBrick-logo-587-BAP.png

Usage and Biology

The Biotinylation Acceptor Peptide (BAP) is a 15 amino acid long peptide identified by Schatz J., 1993 in an library screening approach. This peptide with the sequence 5' - GLNDIFEAQKIEWHE - 3' contains a central lysine that is specifically biotinylated by the prokaryotic enzyme biotin holenzyme synthetase, encoded in the BirA gene of E. coli. Specific biotinylation of this peptide sequence can be performed in vivo by contransfecting a plasmid with the BirA gene as described for the AAV in Arnold et al.; 2006 or by an in vitro coupling approach using the purified Escherichia coli enzyme biotin ligase (BirA).

Restriction sites

Restriction site Enzyme Recognition sequence Activity Temp.
upstream 587 [http://www.neb.com/nebecomm/products/productR3136.asp BamHI-HF] Freiburg10 recognition site BamHI-HF.Gif 100;50;10;100 37°C
downstream 587 [http://www.neb.com/nebecomm/products/productR3151.asp PvuII-HF] Freiburg10 recognition site PvuII-HF.Gif 0;25;0;100 37°C

Sequence and Features


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