Expression constructs for the production of glycosylation enzymes
The Glyco-Enzyme Repository holds the design records for a library of expression constructs covering mammalian glycosyltransferases, glycoside hydrolases and glycan-modifying enzymes, together with a set of bacterial homologues. Every coding region was captured as a Gateway® entry clone and transferred into custom destination vectors for expression in mammalian cells, insect cells (baculovirus) and bacteria. Each record carries the truncation and fusion strategy, the GenBank sequence of every construct, vector maps and the clone ID needed to order the plasmid.
These records document constructs built several decades ago and have not been re-verified against current database releases.
Gene symbols, accessions and annotations are as recorded when each construct was made. Gene nomenclature has since changed for a number of entries — where the Repository knew a symbol under an earlier name, both are shown on the record. Treat the sequence files as authoritative for what is in the plasmid, and re-check external annotation against UniProt before relying on it.
What is in a record
Identification
Gene symbol with any legacy synonym, UniProt accession, NCBI GeneID, DNA and protein RefSeq accessions, the MGC clone used as PCR template, and the CAZy family assignment.
Design strategy
The domain structure and topology of the native enzyme, and the truncation applied to remove transmembrane segments and yield a soluble catalytic domain.
Gateway entry clones
pDONR221 capture clones for N- and C-terminal fusion strategies, available for transfer into user-generated destination vectors.
Expression constructs
Every host/vector combination built for that gene — pGEn1–3 and pGEc1–2 for mammalian cells, polyhedrin-driven baculovirus vectors, and pET16/pET32 for E. coli.
Sequence files
The annotated GenBank sequence of each construct, plus vector maps and full sequence PDFs, served directly from this site.
Ordering information
The DNASU clone ID for each distributable plasmid, with a copy button and a link into the DNASU search, or the contact for baculovirus stocks.
Where to start
- Gene Records — one filterable table of all 386 records. Start here if you know the gene symbol.
- Enzyme Families — browse by CAZy family, or by function for the enzymes CAZy does not classify.
- Search — free-text search across gene symbols, accessions, annotations, vectors and clone IDs.
- Construct Design — the truncation and fusion strategies behind the collection, with the topology diagrams.
- Vectors & Hosts — the destination vectors, their tags, and the host strains and protocols.
- Tutorial — how to find a construct and order it, including how to find a gene symbol from an enzyme activity.
Collection by enzyme class
| Enzyme class | Records | With DNASU clones | Families | Families represented |
|---|---|---|---|---|
| Glycosyltransferase | 200 | 119 | 43 | GT1, GT2, GT3, GT4, GT6, GT7, GT8, GT10, GT11, GT12, GT13, GT14, GT16, GT17, GT18, GT21, GT22, GT23, GT24, GT25, GT27, GT29, GT31, GT32, GT33, GT35, GT39, GT41, GT43, GT47, GT49, GT50, GT54, GT57, GT58, GT59, GT61, GT64, GT65, GT66, GT68, GT76, GT90 |
| Glycoside hydrolase | 77 | 56 | 26 | GH1, GH2, GH13, GH18, GH20, GH22, GH23, GH27, GH29, GH30, GH31, GH33, GH35, GH37, GH38, GH39, GH47, GH56, GH59, GH63, GH65, GH79, GH84, GH85, GH89, GH99 |
| Glycan-modifying enzyme | 62 | 33 | 0 | — |
| Bacterial GT / GH | 47 | 46 | 0 | — |
Largest families
Construct availability
- Plasmid constructs are distributed by DNASU at Arizona State University. 1080 clone IDs are recorded across the collection.
- Baculovirus stocks are supplied by the Jarvis lab (dljarvis@uwyo.edu), not by DNASU.
- Gateway® pDONR entry clones are available for transfer into user-generated destination vectors for custom projects.
- Expression-level summaries for the whole collection are in the summary spreadsheets.