Structural Genomics Consortium Expression Clones
Verified expression of protein domains
The Structural Genomics Consortium (SGC) provides a unique
resource of verified Expression Clones which are suitable
for generating well characterised and functionally active
proteins or protein domains for various purposes.
PLEASE NOTE
The SGC clones cannot currently be ordered via our
website, please see below for details on how
to order.
Product Description
Clone Collection
The Structural Genomics Consortium is a not-for-profit
organisation set up to promote biomedical research by determining
the structures of human proteins with potential therapeutic
importance. Furthermore, the SGC investigates proteins from human
parasites (mainly apicomplexan proteins). Thus the targets are
selected based on their potential as drug targets or involvement in
disease processes. The availability of high resolution structures
constitutes the foundation for structure-based drug design
projects. Technologically, the SGC focuses on
the interaction of proteins with small molecules (ligands,
inhibitors, substrates and co-factors), and on the coverage of
protein families.
The SGC expression clone collection
contains clones from a variety of families. The common denominator
is that they were all expressed and purified from bacteria and were
used for structure determination. The proteins in general do
not correspond to the full-length genes (transcripts) and in
many cases proteins from the SGC collection represent domains of
interest such as catalytic domains and protein-protein interaction
or ligand binding domains with a focus on non-membrane protein
domains.
The Structural Genomics Consortium
operates in the Universities of Oxford and Toronto and at the
Karolinska Institute in Stockholm. The different SGC nodes share
the core technologies but investigate non-overlapping target areas.
The focus of the Oxford node includes protein kinases,
phosphatases (protein tyrosine phosphatases) , oxidoreductases and
other metabolic enzymes, as well as small G-proteins and signal
transduction proteins (RGS, SOCS, 14-3-3, PDZ domains) together
with lysine demethylases and DNA helicases.
Please use the following links to
view the annotated clone lists:
Oxford_SGC_Clones_web
Stockholm_SGC_Clones_web
Methods and Validation
Expression clones were generated by
PCR and ligation-independent cloning (LIC) into one or more of a
set of vectors. The first choice of vector is pNIC28-Bsa4, with the
expression of the cloned gene driven by the T7-LacO system.
Proteins cloned in this vector are fused to an amino-terminal tag
of 23 residues (MHHHHHHSSGVDLGTENLYFQ*SM) including a hexahistidine
(His6) and a TEV-protease cleavage site. Several alternative
expression vectors have been used with selected targets. The
standard hosts for bacterial expression are derivatives of
BL21(DE3) providing tRNAs for rare codons in the host cells.
Specific host strains were created to co-express either accessory
proteins for expression pf particular target proteins. Each clone
in the collection has been verified by the SGC to express the
expected protein from the appropriate bacterial host. The
library and individual clones are supplied in the MACH-1 strain
of E.coli along with BL21(DE3)
expression strains (BL21(DE3)-R3 and BL21(DE3)-R3-pRARE2).
Please find all the technical
information included in the annotated clone lists.
The full materials and methods
(e.g. protein purification protocols) and additional
construct information on a particular protein can be found
on the SGC website (http://www.thesgc.org/structures).
Data Management
The 3D structures for all proteins
encoded by the SGC collection are available from the Protein Data
Bank (PDB) and the SGC website.
iSee (interactive Structurally enhanced
experience)
To make all the information
available to a wide scientific audience, a new tool for
disseminating annotated structural information was created that
also represents an interactive platform allowing for a continuous
update of the annotation by the scientific community. The concept
(denoted iSee) integrates all the information associated with any
given target solved by SGC into a small, self-contained file. The
file not only allows the direct visualisation of text information,
but also offers an interactive visualisation feature fully
integrated to the structural data being presented. Each
of these files (called an iSee datapack), as well as the software
needed to visualise them (ICM-Browser) are available for free
download from SGC website (http://www.thesgc.org/iSee/).
Applications
The SGC clone collection comprises
clones that allow expression of milligram quantities of purified
protein using the host strains and protocols indicated in the
respective structure pages of the SGC website.
These proteins provide a rich
resource for biochemical investigations, including crystallisation
and biophysical characterisation, small-molecule inhibitor screens,
ligand/drug screening and generation of antibodies.
Background Vectors
Click here for further information on the
vectors used in the SGC clone collection and details on how to
purchase these vectors.
Search and Order
The SGC expression clone
collection is not currently searchable via our GenomeCube search
and cannot be ordered online. We are in the process of
integrating the data, however in the meantime you can still order
these clones using a purchase order sent by email or fax to our
customer services at the contact details below.
To place your order:
1. Search for
the clone you require in the databases at the links below and
identify the Construct ID.
Oxford_SGC_Clones_web
Stockholm_SGC_Clones_web
If you have any
problems finding the clone you are looking for please contact us.
2. Use
the Construct ID to place your purchase order.
Price per
single clone £87
Price for
entire library £950
(Prices exclude
VAT and shipping)
For pricing on
multiple clones please contact our customer services for
a quotation.
3. Print
off, read and sign the appropriate Material Transfer Agreement:
US-only
non-US
4. Send your
order and signed Material Transfer Agreement to:
Fax: +44 (0)
115 973 9021
e-mail: sales@sourcebioscience.com
References
Savitsky P, Bray J, Cooper CD,
Marsden BD, Mahajan P, Burgess-Brown NA, Gileadi O. High-throughput production of human proteins for
crystallization: the SGC experience. J Struct Biol.
172(1):3-13. (2010)
Gileadi O, Knapp S, Lee WH, Marsden
BD, Muller S, Niesen FH, Kavanagh KL, Ball LJ, von Delft F, Doyle
DA, Oppermann UC, Sundstrom M. The scientific impact of the Structural Genomics
Consortium: a protein family and ligand-centered approach to
medically-relevant human proteins J Struct Funct Genomics
15(10):1215-26 (2007)