|
|
SERVICE |
DESCRIPTION |
TIME |
| 1
|
Cloning of a gene
of interest into a Bacterial expression vector |
-
Amplification / isolation of the gene of interest out of a customer-supplied
construct and subcloning it into a bacterial expression vector.
-
Verification of the authenticity of the subcloned gene by restriction enzyme
digest and sequencing
-
Maxi-prep of the recombinant vector DNA
|
1-2 weeks |
| 2
|
Generation and
identification of
bacterial colonies
expressing target
proteins |
-
Transformation of recombinant constructs into high-efficiency expression
bacterial strain
-
Mini-induction to over-express the target protein
-
Test for the expression of the recombinant protein by western blot (customer
must provide appropriate antibodies) if desired.
|
1 week |
| 3
|
Large - Scale
Culture |
-
One litre of bacterial culture will be induced with IPTG and harvested for
protein purification
|
2 days |
| 4
|
Purification |
-
Protein purification using Ni-NTA (for 6xHIS-tagged proteins) or glutathione
(for GST-tagged proteins) beads at either native or denatured conditions
|
1 week |
| 5
|
Total |
|
3-4 weeks |
Note:
* If customer does not provide the gene, there will be additional charges (both
cloning and sequencing is included) and it may need 1-2 weeks to amplify the
gene from library and to confirm the sequences.
** We do not guarantee the yield of purified protein since it changes from
protein to protein. More large-scale culture may be needed to obtain a certain
amount of protein for antibody production.
|
