Web: www.ImmunoK.com
Email: IKinfo@amsbio.com
IMMUNOKONTACT CUSTOM SERVICES:
Protein Expression in Bacterial System
1
SERVICE
Cloning of a gene
of interest into a
bacterial
expression vector



DESCRIPTION
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.
TIME
2
weeks
Transformation of recombinant constructs into highefficiency 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
Generation and
identification of
bacterial colonies
expressing target
proteins

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
2


4
weeks
Total
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.
UNITED KINGDOM, INTERNATIONAL:
AMS BIOTECHNOLOGY (EUROPE) LTD.
SWITZERLAND
AMS BIOTECHNOLOGY (EUROPE) LTD.
DEUTSCHLAND:
AMS BIOTECHNOLOGY GMBH
63B MILTON PARK, ABINGDON OXON OX14 4RX
CENTRO NORD-SUD 2E, 6934 BIOGGIO LUGANO
HENKELSTRASSE 15, 65187 W IESBADEN
TEL: +44 (0) 1235 828200
FAX: +44 (0) 1235 820482
TEL: +41 (0) 91 6045522
FAX: +41 (0) 91 6051785
TEL: +49 (0) 69-779099
FAX: +49 (0) 69-13376880
Web: www.ImmunoK.com
Email: IKinfo@amsbio.com
IMMUNOKONTACT CUSTOM SERVICES:
Protein Expression in Baculoviral System
1
2
3
4
5
SERVICE
Cloning of a gene
of interest into a
baculoviral
expression vector
Generation,
identification, and
plaque purification
of baculoviral
colonies expressing
target proteins
Amplification of
recombinant
baculovirus for
high titre stock
Large-scale culture
of recombinant
baculovirus
Purification











DESCRIPTION
Amplification/isolation of the gene of interest out of a
customer-supplied construct and subcloning it into a
baculoviral expression vector.
Verification of the authenticity of the subcloned gene by
restriction enzyme digest and sequencing.
Maxi-prep of the recombinant vector DNA.
Transfection of insect cells (SF9) with the recombinant
transfer vector (gene of interest cloned in the vector) and
baculoviral DNA.
Isolation and plaque-purification of ten baculoviral
recombinant clones.
Test for expression of the recombinant protein by western
blot (customer must provide appropriate antibodies) if
required.
Generation of 500 ml of high-titre viral stock from low-titre
stock solution (approximately 10 ml is required) of a single
recombinant.
A few rounds of amplification of virus may be needed if the
titre of initial virus is too low.
Determination of the titre by end-point dilution assay.
One litre of insect cells will be infected with high-titre virus
stocks (25-30 ml of high titre virus required).
Protein purification using a Ni-NTA (for 6xHIS-tagged
proteins) or glutathione (for GST-tagged proteins) column at
either native or denatured condition.
TIME
1-2
weeks
3
weeks
1 week
1 week
1 week
8
weeks
Total
Note:
*
If customer does not provide the gene, there will be additional charges for both cloning and sequencing 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.
UNITED KINGDOM, INTERNATIONAL:
AMS BIOTECHNOLOGY (EUROPE) LTD.
SWITZERLAND
AMS BIOTECHNOLOGY (EUROPE) LTD.
DEUTSCHLAND:
AMS BIOTECHNOLOGY GMBH
63B MILTON PARK, ABINGDON OXON OX14 4RX
CENTRO NORD-SUD 2E, 6934 BIOGGIO LUGANO
HENKELSTRASSE 15, 65187 W IESBADEN
TEL: +44 (0) 1235 828200
FAX: +44 (0) 1235 820482
TEL: +41 (0) 91 6045522
FAX: +41 (0) 91 6051785
TEL: +49 (0) 69-779099
FAX: +49 (0) 69-13376880