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

Revision as of 21:36, 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 (18 first amino acids) 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

@@ Line 11: / Line 11: @@
 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.
+Moreover we decide to suppres the peptid sigal (18 first amino acids) which is cleaved in order to conserve the fusion.
 ===Source===