G protein alpha-s(alpha3beta5/G226A)
(Plasmid
#55798)
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PurposeIn addition to the mutations in alpha-s(alpha3beta5), this dominant negative G protein alpha-s mutant contains the G226A mutation, which impairs activating conformation changes in Switch II.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 55798 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNAI/Amp
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 4800
- Total vector size (bp) 6387
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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 namealpha-s (alpha3beta5/G226A)
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Alt nameGnas
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SpeciesR. norvegicus (rat)
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Insert Size (bp)1586
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MutationN271K, K274D, R280K, T284D, I285T, G226A in alpha-s sequence
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Entrez GeneGnas (a.k.a. ALEX, G-alpha-8, Gnas1, Gnpas, Nesp55, SCG6)
- Promoter CMV
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Tag
/ Fusion Protein
- internal EE epitope (residues 189-194 in alpha-s sequence are EYMPTE)
Cloning Information
- 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)
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-s(alpha3beta5/G226A) was a gift from Catherine Berlot (Addgene plasmid # 55798 ; http://n2t.net/addgene:55798 ; RRID:Addgene_55798) -
For your References section:
A highly effective dominant negative alpha s construct containing mutations that affect distinct functions inhibits multiple Gs-coupled receptor signaling pathways. Berlot CH. J Biol Chem. 2002 Jun 7;277(23):21080-5. Epub 2002 Apr 1. 10.1074/jbc.M201330200 PubMed 11927592