IGEM:MIT/2005/Receiver Head Unit: scFv

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

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.

Current Status
Experiments Status

FWD for Testing scFvs using anti-FLAG CATCAT GAATTC GCGGCCGC T TCTAGA TG GACGTCGTTATGACTCAAACACC -rand- -EcoRI --NotI--  -XbaI[---]      Primer start


 * 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

 * To test whether the scFvs are expressed? How?
 * We need constitutive promoter with RBS upstream of it. There is one in the library available. Biobrick scFv onto RBS/promoter, and it should express. Do a western blot to see if protein expression by using FLAG tag. We need anti-FLAG antibody (ask Natalie).

../Completed Work/

Additional Information
 Anti-rabbit 
 * Sequence: Sequence of synthetic construct anti-rabbit IgG scFV mRNA, partial cds
 * Paper: scFv Antibody Piezoimmunosensors
 * Summary: ../Summary scFvSensor/