CFP(159-238)-beta-1 in pcDNAI/Amp
(Plasmid #55592)

• Purpose: A carboxyl-terminal CFP fragment was fused to Gbeta-1. When co-expressed with an amino-trerminal CFP or YFP fragment fused to a Ggamma subunit, a fluorescent signal is produced.
• Depositing Lab: Catherine Berlot
• Publication: Mervine et al Mol Pharmacol. 2006 Jul;70(1):194-205. Epub 2006 Apr 26.

Backbone

• Vector backbone: pcDNAI/Amp
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4800
• Total vector size (bp): 6058
• Modifications to backbone: A BglII site was introduced into the polylinker of pcDNA1/Amp 3' to the 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: CFP (159-238)-Beta 1
• Alt name: transducin beta
• Species: H. sapiens (human); Aequorea victoria
• Insert Size (bp): 1258
• Mutation: CFP(159-238) includes a substitution of His for Asn-165. 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 CFP(159-238) destroyed the BamHI site.
• Entrez Gene: GNB1 (a.k.a. HG2A, MDS, MRD42)
• Promoter: CMV
• Tag / Fusion Protein:
  • CFP(159-238) 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: pECFP 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