Available Tagged Genes : Ordering
Complete mouse and human genes that express GFP, luciferase or epitope-tagged proteins.
SpectraGenetics offers full length genes with GFP, RFP, BFP, luciferase or epitope-tag inserts. Each clone contains the full complement of exons and introns, plus thousands of nucleotides upstream and downstream of the transcribed sequence. Unlike cDNA-based constructs, SpectraGenetics clones exhibit native levels of transcription and native pattern of alternative splicing.
Applications include assays for:
- Protein abundance and localization
- Transcriptional and translational regulation
- Post-translational modifications
- Protein-protein interactions
- Alternative splicing
- Knockdown of RNA and protein by RNAi
- Transcription factor binding sites (ChIP)
SpectraGenetics’ tagged genes are based on end-sequenced fosmid clones, not cDNA. Each tagged clone carries the complete gene of interest embedded in approximately 40 kb of native human or mouse genomic DNA, with the reporter sequence (Renilla mulleri GFP, Gaussia princeps luciferase or epitope tag) fused to the extreme 5′- or 3′-end of the protein-coding sequence.
Features of SpectraGenetics’ tagged genes include:
Tagged proteins are produced at native levels. Our tagged genes retain genomic sequences required for natural regulation of transcription, including introns and thousands of nucleotides upstream and downstream of the transcription unit. As a result, tagged proteins are expressed at physiologically correct times and levels. In contrast, proteins expressed from standard cDNA expression vectors are not expressed at physiologically appropriate times or levels.
The full repertoire of splice-isoforms is produced. Because all native introns and exons are present in our tagged genes, physiologically appropriate splicing isoforms are produced. In contrast, standard cDNA expression vectors yield just a single isoform.
Tagged proteins exhibit native subcellular localization and native biological function. Proteins carrying reporter tags at their termini have a high likelihood of exhibiting native biological function. And, because they are expressed under the same regulatory controls as their untagged counterparts, they generally exhibit appropriate subcellular localization and do not perturb the natural physiology of the cell. In contrast, proteins that are over-expressed from standard expression vectors are more likely to mislocalize and/or perturb normal cell physiology.
GFP-tagged proteins carry Renilla mulleri GFP, a fluorescent protein that is as bright or brighter than other green fluorescent proteins including EGFP. The excitation and emission spectra of Renilla GFP are compatible with standard fluorescein or GFP filter sets.
Luciferase-tagged proteins carry Gaussia princeps luciferase , an ATP-independent luciferase that is as bright or brighter than other known luciferases.
Clones are available with neomycin or puromycin resistance genes in the fosmid backbone to facilitate production of stable transfectants.
A research-use license to the tagged gene accompanies each order.