GFP-C1-PLCdelta-PH
(Plasmid #21179)

• Purpose: encodes a PI(4,5)P2 lipid selective PH domain that can be used as a fluorescent translocation biosensor to monitor changes or local differences in the concentration of plasma membrane PI(4,5)P2 lipids
• Depositing Lab: Tobias Meyer
• Publication: Stauffer et al Curr Biol. 1998 Mar 12. 8(6):343-6.

Backbone

• Vector backbone: C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: PLCdelta
• Species: H. sapiens (human)
• Insert Size (bp): 529
• Mutation: PH domain of human PLC delta, amino acids Met1–Ile175. (Lab plasmid WC 0003)
• Entrez Gene: PLCD1 (a.k.a. NDNC3, PLC-III)
• Tag / Fusion Protein:
  • GFP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (unknown if destroyed)
• 3′ cloning site: XbaI (unknown if destroyed)
• 5′ sequencing primer: n/a

Resource Information

• Articles Citing this Plasmid:
  • 74 References

Terms and Licenses

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

Depositor Comments

This construct encodes a PI(4,5)P2 lipid selective PH domain that can be used as a fluorescent translocation biosensor to monitor changes or local differences in the concentration of plasma membrane PI(4,5)P2 lipids (Stauffer TP, Ahn S, Meyer T; Curr Biol. 1998 8:343-6). It is best used in combination with a second fluorescent protein that uniformly marks the plasma membrane (the last 20 amino acids "CAAX" region of KRas is often used as a PM marker). Relative changes in cell-wide or local plasma membrane PI(4,5)P2 can be determined by using a mask for the PM region and by dividing the two background subtracted PH domain and CAAX images. At high expression levels, this construct interferes with PI(4,5)P2 signaling and alters cortical actin (Raucher D et al Cell. 2000 100:221-8).