High throughput protein production and crystallization at NYSGXRC.

Michael J. Sauder, Marc E. Rutter, Kevin Bain, Isabelle Rooney, Tarun Gheyi, Shane Atwell, Devon A. Thompson, Spencer Emtage, Stephen K. Burley

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

Phase II of the Protein Structure Initiative, funded by the NIH NIGMS (National Institute of General Medical Sciences), is a 5-year effort to determine thousands of protein structures. The New York SGX Research Center for Structural Genomics (NYSGXRC) is one of the four large-scale production centers tasked with determining 100-200 structures annually. Almost all protein production is carried out using the high throughput structural biology platform at SGX Pharmaceuticals (SGX), which supplies 120 or more ultrapure proteins per month for NYSGXRC crystallization and structure determination activities. Protocols for PCR, cloning, expression/solubility testing, fermentation, purification, and crystallization are described. General protocols and detailed experimental results for each target are updated weekly at the public PepcDB website (pepcdb.pdb.org/), and all NYSGXRC clones should be available in 2008 through the PlasmID resource operated by the Harvard Institute of Proteomics.

Original languageEnglish (US)
Pages (from-to)561-575
Number of pages15
JournalMethods in molecular biology (Clifton, N.J.)
Volume426
DOIs
StatePublished - 2008

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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