Difference between revisions of "Part:BBa J32006:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J32006 short</partinfo>
 
<partinfo>BBa_J32006 short</partinfo>
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===Design Notes===
 
===Design Notes===
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Fusion Protein
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The '''pelB''' leader sequence is a sequence of amino acids which when attached to a protein, directs the protein to the periplasmic membrane of E. coli, where the sequence is removed by pelB peptidase. It is used to direct coat protein-antigen fusions to the cell surface.
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 +
The '''antibody''' is currently marked as a cut site between Pel B and S*Tag.  When the antibody sequence is determined it will follow this cut site.
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The '''S•Tag''' sequence is a novel fusion peptide tag for recombinant proteins that allows detection by a rapid, sensitive homogeneous assay or by colorimetric detection in Western blots. Proteins can also be rapidly purified from crude extracts using a one step affinity separation method. The system is based on the strong interaction between the 15aa S•Tag and 103aa S-protein.
  
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Neisseria gonorrhoeae '''IgA Protease''' NCBI Accession AE004969.  Allows for suface display of antibodies.
  
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It is a fusion protein so biobrick prefixes and suffixes have been modified to 10 and 24 bp, removing the bp between the flanking restriction sites and the sequence. Otherwise it would read out of frame with 8bp overlap.
  
 
===Source===
 
===Source===
  
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The signal sequence PelB and S*Tag sequence came from Novagen. The antibody sequence is for cAb-CEA5 nanobody from Ablynx. The C-terminus of Neisseria gonorrhoeae IgA Protease NCBI accession number AE004969
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The signal sequence PelB and S*Tag sequence came from Novagen. The C-terminus of Neisseria gonorrhoeae IgA Protease NCBI accession number AE004969
  
 
===References===
 
===References===

Latest revision as of 04:17, 12 July 2006

Display


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1440
    Illegal PstI site found at 1566
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 364
    Illegal PstI site found at 1440
    Illegal PstI site found at 1566
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 638
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1440
    Illegal PstI site found at 1566
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1440
    Illegal PstI site found at 1566
    Illegal NgoMIV site found at 54
    Illegal NgoMIV site found at 649
    Illegal NgoMIV site found at 1274
    Illegal AgeI site found at 1049
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 119
    Illegal BsaI.rc site found at 330
    Illegal SapI.rc site found at 530


Design Notes

The pelB leader sequence is a sequence of amino acids which when attached to a protein, directs the protein to the periplasmic membrane of E. coli, where the sequence is removed by pelB peptidase. It is used to direct coat protein-antigen fusions to the cell surface.

The antibody is currently marked as a cut site between Pel B and S*Tag. When the antibody sequence is determined it will follow this cut site.

The S•Tag sequence is a novel fusion peptide tag for recombinant proteins that allows detection by a rapid, sensitive homogeneous assay or by colorimetric detection in Western blots. Proteins can also be rapidly purified from crude extracts using a one step affinity separation method. The system is based on the strong interaction between the 15aa S•Tag and 103aa S-protein.

Neisseria gonorrhoeae IgA Protease NCBI Accession AE004969. Allows for suface display of antibodies.

It is a fusion protein so biobrick prefixes and suffixes have been modified to 10 and 24 bp, removing the bp between the flanking restriction sites and the sequence. Otherwise it would read out of frame with 8bp overlap.

Source

The signal sequence PelB and S*Tag sequence came from Novagen. The C-terminus of Neisseria gonorrhoeae IgA Protease NCBI accession number AE004969

References