IGEM:MIT/2005/Receiver Head Unit: scFv
Location(s): BUG Lounge, Kate's Lab
- 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.
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
- 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).