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

m
m
Line 6: Line 6:
  
 
===Design Notes===
 
===Design Notes===
*In our project we use g3p as a fusion to OmpA-Linker (<partinfo>Part:BBa_K103996</partinfo>) which need SacI restriction site for inframe fusion.
+
*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.
 
*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 signal peptide (18 first amino acids) which is cleaved in order to conserve the N-ter fusion.
 
*Moreover we decide to suppres the signal peptide (18 first amino acids) which is cleaved in order to conserve the N-ter fusion.

Revision as of 14:38, 30 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 signal peptide (18 first amino acids) which is cleaved in order to conserve the N-ter 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