YFP(1-158)-beta-1 in pcDNAI/Amp
(Plasmid #54468)

• Purpose: An amino-terminal YFP fragment was fused to Gbeta-1. When co-expressed with a carboxyl-terminal YFP or CFP fragment fused to a Ggamma subunit, a fluorescent signal is produced.
• Depositing Lab: Catherine Berlot
• Publication: Hynes et al J Biol Chem. 2004 Jul 16;279(29):30279-86. Epub 2004 May 10.

Backbone

• Vector backbone: pcDNAI/Amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 6283
• Modifications to backbone: BglII introduced to polylinker of pcDNA1/amp 3' of BamHI site.
• 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: YFP (1-158)/beta-1
• Alt name: transducin beta
• Species: H. sapiens (human); Aequorea victoria
• Insert Size (bp): 1483
• Mutation: GBeta-1 was amplified via PCR, which removed an internal BglII site and added an N-terminal linker sequence (RSIAT), a 5' BamHI site, and a 3' BglII site. Cloning into the BglII site of YFP(1-158) destroyed the BamHI site. PCR amplification of YFP (1-158) introduced a substitution of M for Q69.
• Entrez Gene: GNB1 (a.k.a. HG2A, MDS, MRD42)
• Promoter: CMV
• Tag / Fusion Protein:
  • YFP(1-158) was fused to the amino terminus of Gbeta1. (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: BglII (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 purchased from Clontech. beta-1 cDNA obtained from Janet Robishaw (Weis Center for Research, Geisinger Clinic, Danville PA).

Terms and Licenses

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