Difference between revisions of "Part:BBa K257001:Design"

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===Design Notes===
 
===Design Notes===
  
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In our project we use g3p as a fusion to OmpA-Linker (BBa_K103996) which need SacI restriction site for inframe fusion.
  
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The g3p sequenced comes from the phage genome.
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So we design g3p with SacI site at the N-terminal. SacI (GAGCT^C) site is shared with XbaI (T^CTAGA) in order to have SacI site for fusion and standard sites.
  
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It's a protein involved in the adhesion of the phage to the bacteria.
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Moreover we decide to suppres the peptid sigal which is cleaved in order to conserve the fusion.
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Its sequence is surounded by the biobrick enzymatic restriction sites.
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===Source===
 
===Source===

Revision as of 21:35, 26 August 2009

Gene 3 protein of filamentous phages without the 18 first amino acids


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 592
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In our project we use g3p as a fusion to OmpA-Linker (BBa_K103996) which need SacI restriction site for inframe fusion.

So we design g3p with SacI site at the N-terminal. SacI (GAGCT^C) site is shared with XbaI (T^CTAGA) in order to have SacI site for fusion and standard sites.

Moreover we decide to suppres the peptid sigal which is cleaved in order to conserve the fusion.

Source

g3p could be found in filamentous bacteriophages like M13, fd, f1, etc... or in phage helper like M13KO7, etc...

References

  • The Mechanism of Bacterial Infection by Filamentous Phages Involves Molecular Interactions between TolA and Phage Protein 3 Domains. Fredrik Karlsson, Carl A. K. Borrebaeck, Nina Nilsson, and Ann-Christin Malmborg-Hager
  • Interdomain interactions within the gene 3 protein of philamentous phage. Jean Chatellier, Oliver Hartley, Andrew D. Grifths, Alan R. Fershta, Greg Wintera, Lutz Riechmannb
  • A prokaryotic membrane anchor sequence: Carboxyl terminus of bacteriophage fl gene III protein retains it in the membrane. Jef D. Boeke AND Peter Model