SGC expression clones

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)