In order to perform large-scale analysis of protein complexes in Escherichia coli, researchers at the University of Toronto have created a library of affinity-tagged open reading frames. With c-terminal tandem affinity purification (TAP) or sequential peptide affinity (SPA) tagged alleles of over 800 ORFs, the E. coli Tagged ORF library allows purification of tagged proteins and their interacting protein partners.
To create this library, ORFs were selected to represent broad biological coverage, including highly-conserved essential and non-essential proteins, proteins with putative functional assignments, and hypothetical uncharacterized ORFs. Then, using a phage recombination system, sequence-specific PCR products encoding affinity purification tags were inserted into discrete loci of the E. coli chromosome. 857 proteins, including 198 essential and conserved proteins, were successfully tagged. Over a quarter of these proteins were also successfully purified by the source lab. Proteins were isolated by affinity purification and their binding partners identified using LC-MS and MALDI-TOF mass spectrometry.
- Generate a reliable network of functionally diverse protein complexes
- Gain insight into the function of uncharacterized proteins
- Outline the topological organization of the bacterial interactome
We provide certain clone resources developed by leading academic laboratories. Many of these resources address the needs of specialized research communities not served by other commercial entities. In order to provide these as a public resource, we depend on the contributing academic laboratories for quality control.
Therefore, these are distributed in the format provided by the contributing institution "as is" with no additional product validation or guarantee. We are not responsible for any errors or performance issues. Additional information can be found in the product manual as well as in associated published articles (if available). Alternatively, the source academic institution can be contacted directly for troubleshooting.
Clones are provided as a live culture in a 2 mL tube. Each tube contains LB medium supplemented with 8% glycerol and the appropriate antibiotic. We will ship your clone at room temperature via express delivery. Store the stock clone at -80°C.
Orders of 50 clones or greater and stock plates will be provided in microtiter plates. These will ship on dry-ice via overnight delivery and should be stored at -80°C immediately upon receipt. Please contact our customer service department for a quote and estimated shipping time on bulk orders.
Orders for the entire C. elegans Promoter collection are provided in 96-well microtiter plates. These will ship on dry-ice via overnight delivery and should be stored at -80°C immediately upon receipt. Please contact our customer service department for estimated shipping time on orders for the entire collection.
- G. Butland et al.,Interaction network containing conserved and essential protein complexes in Escherichia coli. Nature. 433, 531–537 (3 February 2005).
- M. Zeghouf et al.,Sequential peptide affinity (SPA) system for the identification of mammalian and bacterial protein complexes. Journal of Proteome Research. 3, 463–468 (2004).
- G. Rigaut et al., A generic protein purification method for protein complex characterization and proteome exploration. Nat. Biotechnol. 17(10), 1030-1032 (October 1999).