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PurposeThis G protein alpha-q construct contains internal insertions of GFP and the EE epitope
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 66080 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA1/Amp
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 4800
- Total vector size (bp) 7119
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Vector typeMammalian Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH10B
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameG protein alpha-q-EE-YFP
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Alt nameGnaq
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SpeciesM. musculus (mouse)
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Insert Size (bp)2319
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MutationBamHI and SacI sites in alpha-q were removed with silent mutations.
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Entrez GeneGnaq (a.k.a. 1110005L02Rik, 6230401I02Rik, Dsk1, Dsk10, Galphaq, Gq, GqI)
- Promoter CMV
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Tags
/ Fusion Proteins
- GFP was inserted internally into the alpha-q sequence between residues 124 and 125 with a SGGGGS linker on each end of the YFP.
- The EE epitope was introduced into the alpha-q sequence by mutating residues 171-176 (SYLPTQ) to EYMPTE.
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NotI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer SP6 (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made bypEGFP was purchased from Clontech.
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Article Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- 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.
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For your Materials & Methods section:
G protein alpha-q-GFP was a gift from Catherine Berlot (Addgene plasmid # 66080 ; http://n2t.net/addgene:66080 ; RRID:Addgene_66080) -
For your References section:
Visualization of a functional Galpha q-green fluorescent protein fusion in living cells. Association with the plasma membrane is disrupted by mutational activation and by elimination of palmitoylation sites, but not be activation mediated by receptors or AlF4-. Hughes TE, Zhang H, Logothetis DE, Berlot CH. J Biol Chem. 2001 Feb 9;276(6):4227-35. Epub 2000 Nov 13. 10.1074/jbc.M007608200 PubMed 11076942