IGEM:MIT/2005/Receiver Head Unit: scFv

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Revision as of 07:04, 19 July 2005 by >Jenny (→‎Experiments)
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POC

Jenny

Function

  • To provide a landing site for ligand.
  • To cause dimerization/conformational change in the Receiver.
  • To transmit signal to Receiver.
  • Will use anti-fluorescein scFv for test phase.
  • Ultimately use anti-rabbit scFv for system.

Device Depiction

Device Parts

Current Status

  • Primers have been designed for biobrick format.
  • pCT-4-4-20, pCT302 S101A, pCT-4M5.3 anti-fluorescein plasmids miniprepped
FWD: 5' CATCAT GAATTC GCGGCCGC T TCTAGA GACGTCGTTATGACTCAAACACC 3'
         rand   EcoRI   NotI      XbaI       Primer
REV: 5' GCTCAT CTGCAG GCGGCCGC T ACTAGT TTATTA CTTATCATCATCATCCTTATAATC GGAGACGGTGACTGAGGTTCC 3'
         rand   PstI   NotI       SpeI   Stop          FLAG Tag                 Primer

Experiments

  • express scfv as single protein, test with biotin / streptaviden??
  1. Test for tags that surround scFv sequence
    • Flag tag on 3' end
      • Use antibodies that recognize those tags.
      • Western, indirect immunofluorescence, permiabilize

Open Issues/Questions

Additional Information

Anti-rabbit

Sequence: Sequence of synthetic construct anti-rabbit IgG scFV mRNA, partial cds
Paper: scFv Antibody Piezoimmunosensors
Summary: [[../Summary scFvSensor/]]