G protein alpha-o-YFP
(Plasmid #55776)

• Purpose: This G protein alpha-o construct contains internal insertions of YFP and the EE epitope
• Depositing Lab: Catherine Berlot
• Publication: Yost et al Mol Pharmacol. 2007 Oct;72(4):812-25. Epub 2007 Jun 27.

Backbone

• Vector backbone: pcDNA1/amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 7639
• 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-o internally tagged with EYFP and EE epitope
• Alt name: Gnao
• Species: R. norvegicus (rat)
• Insert Size (bp): 2838
• Mutation: A 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 Gene: Gnao1 (a.k.a. Gnao, Hg1g, RATBPGTPC)
• Promoter: CMV
• Tags / 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 Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: SP6

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pEYFP was purchased from Clontech.

Terms and Licenses

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