G protein alpha-o-YFP
PurposeThis G protein alpha-o construct contains internal insertions of YFP and the EE epitope
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||55776||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4800
- Total vector size (bp) 7639
Vector typeMammalian Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Copy numberHigh Copy
Gene/Insert nameG protein alpha-o internally tagged with EYFP and EE epitope
SpeciesR. norvegicus (rat)
Insert Size (bp)2838
MutationA BglII site was removed from the 5' untranslated region of alpha-oEE and then a unique BglII site was introduced in frame between Pro119 and Phe120 into which EYFP containing a substitution of Met for Gln69 and flanked on both ends by a SGGGGS linker was inserted as a BamHI cassette (destroying both the BglII and BamHI sites).
Entrez GeneGnao1 (a.k.a. Gnao, Hg1g, RATBPGTPC)
- Promoter CMV
/ Fusion Proteins
- EYFP containing a Q69M mutation and flanked on both ends by a SGGGGS linker was inserted between Pro119 and Phe120.
- EE epitope was introduced internally with D167E and Q169M mutations.
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer SP6 (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:G protein alpha-o-YFP was a gift from Catherine Berlot (Addgene plasmid # 55776 ; http://n2t.net/addgene:55776 ; RRID:Addgene_55776)
For your References section:Live cell analysis of G protein beta5 complex formation, function, and targeting. Yost EA, Mervine SM, Sabo JL, Hynes TR, Berlot CH. Mol Pharmacol. 2007 Oct;72(4):812-25. Epub 2007 Jun 27. 10.1124/mol.107.038075 PubMed 17596375