Matrix-roGFP
(Plasmid #49437)

• Purpose: Expresses thiol redox-sensitive ratiometric sensor roGFP in the mitochondrial matrix of mammalian cells
• Depositing Lab: Paul Schumacker
• Publication: Waypa et al Circ Res. 2010 Feb 19;106(3):526-35. doi: 10.1161/CIRCRESAHA.109.206334. Epub 2009 Dec 17.

Backbone

• Vector backbone: VQ Ad5CMV K-NpA
• Backbone manufacturer: ViraQuest Inc.
• Backbone size w/o insert (bp): 6217
• Total vector size (bp): 6970
• Modifications to backbone: None
• Vector type: Mammalian Expression, Adenoviral

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: Matrix-roGFP2
• Alt name: Mitochondrial roGFP2
• Species: Synthetic
• Insert Size (bp): 780
• Mutation: C48S, Q80R, S147C, and Q204C in Clontech's GFP
• Promoter: CMV
• Tag / Fusion Protein:
  • Cytochrome c oxidase subunit IV mitochondrial targeting sequence (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: 5'GGCGTGTACGGTGGGAGGTCTATA-3'
• 3′ sequencing primer: 5'-TTAAAAAACCTCCCCACCTCCCCC-3'

Resource Information

• Supplemental Documents:
  • Description of Matrix-roGFP
• A portion of this plasmid was derived from a plasmid made by: We inserted mutations into Clontech's eGFP gene to convert it into roGFP and cloned by PCR using a primer to insert the mitochondrial targeting sequence for cyt c oxidase subunit IV 5' to the roGFP2. This entire construct was then ligated into the VQ vector between KpnI and NotI sites.
• Articles Citing this Plasmid:
  • 20 References

Terms and Licenses

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