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Team:UC Davis/Adding secretion
From 2009.igem.org
The purpose of the
secretion system is to
introduce a method of secreting target proteins we wish to synthesize
in the
marvelous host: E.coli. For our secretion system we have taken the idea
from (15). Park et.
al showed
that construct containing the truncated from of ice nucleation
protein (INPNC) where found to complete coding region of phaZ1,
including its
signal sequence could lead to stable secretion of the enzyme (15). We have
decided to see if we could co-opt this system to make a more
generalized
inducible system for protein secretion in which the INPNC and phaZ1
signal
sequence serve as carries for a target protein.
Possible challenges:
1. Efficient recognition of the change site may
()
some of the phaZ1 protein we have only cloned the
signal sequence up to the cleavage site.
2. The system may only work for protein in a narrow site range.
3. The expression levels of the system may be low.
Advantages:
- Testing multiple
different combinations, Signal Sequence plus ompA/INPNC would help us
find the best combination for our secretion system based on their
secretion ability and rank them from strongest to weakest in strength
(in this specific system).
- Also, we have
submitted the BioBricks to the parts registry; therefore they can be
used in future studies (for different purposes).
Secretion Models:
Click on a specific model for more information:
There
is wide range of genes present in our environment. Therefore, it is
important
to measure our secretion system’s ability to secrete genes of various
sizes.
| Name |
Molecular
Weight (kD) |
| Myosin |
200.0 |
| -Galactosidase |
116.3 |
| Phosphorylase
b |
97.4 |
| Ovalbumin |
45.0 |
| Carbonic
anhydrase |
31.0 |
| Aprotinin |
6.5 |
| Insulin,
B chain, oxidized |
3.5 |
