Expression constructs for the production of human glycosylation enzymes

Goals
  • Create expression reagents and protocols to produce recombinant human glycosylation enzymes. 
  • Provide enzymes for chemoenzymatic glycan synthesis as standards, recombinant glycoprotein remodeling, synthesis of rare unique glycans
  • Advance understanding of mammalian glycosylation enzymes and glycan synthesis.
Milestones
  • Expression constructs encoding ALL mammalian glycosylation enzymes generated
    • Glycosyltransferases (GTs), glycoside hydrolases (GHs), and glycan modifying enzymes.
  • Production as secreted soluble enzymes catalytic domains (when possible):
    • Truncate transmembrane domains, replace with signal sequences and fusion tags or other larger fusion proteins to facilitate detection, quantitation, and affinity purification.
  • Transfer to expression vectors for production in mammalian cells, insect cells (baculovirus), and bacteria
  • Provide protocols for protein production in each host system.
Construct Availability Navigating this site
  • All constructs are available from DNASU (includes >800 mammalian expression constructs in multiple vectors
  • Baculovirus stocks (>250) are available from the Jarvis lab.
  • Gateway® pDONR constructs are available for transfer into user-generated Gateway destination vectors for custom projects.
  • Design criteria, construct sequences, and clone IDs are available in this website with links to acquiring the clones from the PSI Materials Repository at DNASU and baculovirus stocks from the Jarvis lab.
  • SEE SPREADSHEAT SUMMARY of expression data for all expression constructs!!!
  • Left menubar contains links for navigation to design strategies, gene records, and links to order clones from DNASU.
  • Constructs can be searched by alphabetical list or by CAZy family.
  • A TUTORIAL for navigation of this site is available.
  • Constructs are designated by gene symbol (gene name) rather than enzyme nomenclature (enzyme nomenclature can be VERY ambiguous).
  • Gene symbols (gene names) can be found by searching UniProt or HUGO Gene Nomenclature Committee search engines.