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Team:UC Davis/Adding secretion/model 5
From 2009.igem.org
Secretion Model 5:
Click on an individual part for more information.
a.
INPNC:Ice-nucleation protein (INP) from Pseudomonas Syringae was suggested to be used for display of foreign proteins on the surface of E. coli(7).Furthermore, researches have shown that an INP derivative constituting the N-and C-terminal domains can and has been used to display foreign proteins on the surface of E. coli(9). In our project we are intending to harness and make use of this feature by fusing a specific protein to it.
We
have
modified this protein to Biobrick standard, Tom Knights Standard.
For more information go to: BBa_K265008
OmpA: OmpA
is one of the proteins on the outer
membrane of E. coli (13). OmpA has been found to be useful as
utilizable
fusion part that can fuse our protein to and display on the surface of E.
coli. This part has already been documented on the parts registry;
however,
it has not been tested via fusion with a target protein linked with a
cleavable
signal sequence.
We
have
modified this protein to Biobrick standard, Tom Knights Standard.
Note:
“It has
remained essentially unknown how proteins of E. coli outer membrane are
sorted
and incorporated into this membrane” (10)
For
more
information go to:
BBa_K103006
RBS:
Ribosome
Binding site number 32 (BBa_J61132)
from the registry is being used in our secretion system.
For
more
information go to: BBa_J61132
Terminator: We
are using BBa_B0015, a double terminator, as
our terminator in both our secretion and pH system.
For
more
information go to: BBa_B0015
GFP: We
are using Green Fluorescent Protein as a
reporter that also serves as a small protein in testing our secretion
system.
For more informaiton go to: BBa_K265003
Luciferase: Luciferase
is a firefly protein that also
fluoresces, so it serves as a reporter as well as a testable large
protein.
For more information go to: BBa_1712019
LacI: One
inducible Promoter which was found in the
part registry.
For
more information go to:
BBa_R0010
SS:This
signal sequence, when placed between
INPNC, contains a cleavable site that allows the target fusion protein
to
‘secrete’ from INPNC. We will do the same with OmpA.
We
have
modified this protein to Biobrick standard, Tom Knights Standard.
For more infromation go to :BBa_K265002
Note: We are using this tag, just in case if the GFP or Luciferase does not work under a plate reader.
