IGEM:MIT/2005/Receiver Head Unit: scFv

POC

Alias: Jenny Location(s): BUG Lounge, Kate's Lab E-mail: JennyN@mit.edu

[[../Experiments#Experiment Status/|This is what I'm working on currently.]]

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

• 

  scFv

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

Additional Information

Anti-rabbit

Sequence: Sequence of synthetic construct anti-rabbit IgG scFV mRNA, partial cds

Paper: scFv Antibody Piezoimmunosensors

Summary: [[../Summary scFvSensor/]]