G protein alpha-s-mCherry
PurposeG protein alpha-s internally tagged with mCherry and EE epitope
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||66968||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4800
- Total vector size (bp) 7094
Vector typeMammalian Expression
Growth in Bacteria
Copy numberHigh Copy
SpeciesR. norvegicus (rat); Discosoma sp.
Insert Size (bp)2284
MutationBamHI sites in the alpha-s cDNA were removed and alpha-s residues 73-84 were replaced with Gly-Ser adding a unique BamHI site into which mCherry was subcloned.
Entrez GeneGnas (a.k.a. ALEX, G-alpha-8, Gnas1, Gnpas, Nesp55, SCG6)
- Promoter CMV
/ Fusion Proteins
- mCherry flanked by SGGGS on each side was inserted in place of alpha-s residues 73- 84.
- internal EE epitope (residues 189-194 in alpha- s sequence are EYMPTE)
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer SP6 (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byAlpha-s-EE was produced in the laboratory of Henry Bourne, University of California, San Francisco, CA. mCherry was obtained from Roger Tsien, University of California, San Diego, CA.
Terms and Licenses
- Not Available to Industry
Reference for alpha-s-EE:
Wilson PT, Bourne HR. Fatty acylation of αz. Effects of palmitoylation and myristoylation on αz signaling. J Biol Chem. 1995;270:9667-75.
Reference for mCherry:
Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien RY. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol. 2004;22(12):1567-72. PubMed PMID: 15558047.
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-s-mCherry was a gift from Catherine Berlot (Addgene plasmid # 66968 ; http://n2t.net/addgene:66968 ; RRID:Addgene_66968)
For your References section:Live cell imaging of Gs and the beta2-adrenergic receptor demonstrates that both alphas and beta1gamma7 internalize upon stimulation and exhibit similar trafficking patterns that differ from that of the beta2-adrenergic receptor. Hynes TR, Mervine SM, Yost EA, Sabo JL, Berlot CH. J Biol Chem. 2004 Oct 15;279(42):44101-12. Epub 2004 Aug 5. 10.1074/jbc.M405151200 PubMed 15297467