XE240 Fz1-ER-GFP
(Plasmid #16815)

• Depositing Lab: Randall Moon
• Publication: Kaykas et al Nat Cell Biol. 2004 Jan . 6(1):52-8.

Backbone

• Vector backbone: pCS2+
• Backbone size w/o insert (bp): 4095
• Vector type: Xenopus Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Fz1-ER-GFP
• Alt name: Fz1
• Species: R. norvegicus (rat)
• Mutation: F36M in FKBP
• Tag / Fusion Protein:
  • gfp

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: SP6

Terms and Licenses

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

Depositor Comments

This is a fusion protein consisting of a FKBP domain at the amino terminus of rat Frizzled 1, and GFP at the C terminus.
The expression of the protein is under the control of a CMV promoter. Once expressed the fusion protein enters the endoplasmic reticulum, where it remains trapped (and acts as a dominant negative Frizzled by trapping endogenous Frizzleds) unless one adds proprietary Ariad small molecule reagents that bind to and unfold the FKBP domain, allowing proteolysis and then processing and membrane targeting of the Frizzled 1. Note that the Frizzleds interact in a homo and heteromeric manner.

Reference: A. Kaykas et al., 2004, Nature Cell Biology vol. 6, pp. 52-58.