G protein alpha-s(alpha3beta5)
(Plasmid #55799)

• Purpose: This G protein alpha-s mutant exhibits increased affinity for and decreased ability to be activated by Gs-protein-coupled receptors as well as decreased ability to activate adenylyl cyclase.
• Depositing Lab: Catherine Berlot
• Publication: Berlot et al J Biol Chem. 2002 Jun 7;277(23):21080-5. Epub 2002 Apr 1.

Backbone

• Vector backbone: pcDNAI/Amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 6373
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: G protein alpha-s with alpha-i2 substitutions in alpha3/beta5 loop
• Alt name: Gnas
• Species: R. norvegicus (rat)
• Insert Size (bp): 1572
• Mutation: N271K, K274D, R280K, T284D, I285T in alpha-s sequence
• Entrez Gene: Gnas (a.k.a. ALEX, G-alpha-8, Gnas1, Gnpas, Nesp55, SCG6)
• Promoter: CMV
• 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: EcoRV (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: SP6

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry