EcMBP165-cpGFP.PPYF.T203V.pRSET
(Plasmid #33373)

• Purpose: a single-wavelength maltose sensor constructed from E. coli MalE and cpGFP
• Depositing Lab: Loren Looger
• Publication: Marvin et al Proteins. 2011 Nov;79(11):3025-36. doi: 10.1002/prot.23118.

Backbone

• Vector backbone: pRSET-A
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2300
• Vector type: Bacterial 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: Improved MBP based Maltose Biosensor
• Species: E. coli
• Mutation: cpGFP146 variant from GCaMP2 inserted between AA165-166 of MBP with T203V mutation in GFP.
• Promoter: T7
• Tags / Fusion Proteins:
  • 6xHIS (N terminal on backbone)
  • 6xHIS (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: none (unknown if destroyed)
• 3′ cloning site: none (unknown if destroyed)
• 5′ sequencing primer: T7 forward

Resource Information

• Supplemental Documents:
  • pMBP165.PPYF.T203V.pRSET.gb
  • pMBP165.PPYF.T203V.pRSET.pdf
  • pMBP165.PPYF.T203V.pRSET.sbd

Terms and Licenses

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

Depositor Comments

The GFP-T203V mutation decreases the fluorescence emission of the apo state and increases that of the saturated state in GCaMP2. In the context of MBP165-cpGFP.PPYF, the T203V mutation decreases the fluorescence emission of the apo state by half, whereas saturated fluorescence and affinity are unchanged, increasing ΔF/F to 6.5.