pFRET-HSP33 cys
(Plasmid #16076)

• Depositing Lab: Ulrich Hammerling
• Publication: Guzy et al Cell Metab. 2005 Jun;1(6):401-8. doi: 10.1016/j.cmet.2005.05.001.

Backbone

• Vector backbone: pECFP-N1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 5190
• 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: High Copy

Gene/Insert

• Gene/Insert name: HSP33 cys
• Species: E. coli
• Insert Size (bp): 900
• Tags / Fusion Proteins:
  • EYFP (N terminal on insert)
  • ECFP (C terminal on backbone)

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Dr. Ursula Jakob, Univ of MI
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Depositor Comments

The HSP-FRET probe was generated by inserting YFP into the pECFP-N1 plasmid between the NheI and BglII sites and then ligating the redox-sensitive regulatory domain from the Escherichia coli HSP-33 between YFP and CFP via the EcoRI and BamHI sites. HSP-33: 210 amino acids; BA000007 region 4241962-4242826. Made in Hammerling lab; used in Waypa et al., Increases in mitochondrial reactive oxygen species trigger hypoxia-induced calcium responses in pulmonary artery smooth muscle cells, Circ Res. 2006 Oct 27; 99(9):970-8 PMID: 17008601.