Production of recombinant G protein-coupled receptor in yeast for structural and functional analysis

Richard A.J. Darby*, Mohammed Jamshad, Ljuban Grgic, William J. Holmes, Roslyn M. Bill

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Original languageEnglish
Title of host publicationG protein-coupled receptors
Subtitle of host publicationessential methods
EditorsDavid R. Poyner, Mark Wheatley
PublisherWiley-Blackwell
Pages85-110
Number of pages26
ISBN (Electronic)978-0-470-74921-0
ISBN (Print)978-0-470-74914-2
DOIs
Publication statusPublished - 2010

Keywords

  • analytical ultracentrifugation (AUC), technique characterizing solution state of macromolecules
  • DNA sequence encoding GPCR of interest - amplified via polymerase chain reaction (PCR)
  • G protein-coupled receptors (GPCRs) - naturally present in native membranes
  • PCR amplification and cloning
  • preparation of expression plasmids
  • protocol 5.2 yeast transformations
  • protocol 5.3 screening for GPCR production in P. pastoris clones
  • pProtocol 5.5 GPCR solubilization and purification
  • recombinant G protein-coupled receptor production in yeast - structural and functional analysis
  • solubilization and purification

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