IGEM:MIT/2005/Receiver Head Unit: scFv: Difference between revisions
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>Jenny |
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==POC== | ==POC== | ||
Jenny | Alias: Jenny<BR> | ||
Location(s): BUG Lounge, Kate's Lab<BR> | |||
E-mail: JennyN@mit.edu<BR><BR> | |||
[[../Experiments#Experiment Status/|This is what I'm working on currently.]] | |||
==Function== | ==Function== | ||
*To provide a landing site for ligand. | *To provide a landing site for ligand. | ||
*To cause dimerization/conformational change in the Receiver. | *To cause dimerization/conformational change in the Receiver. | ||
*To transmit signal to Receiver. | *To transmit signal to Receiver. | ||
*Will use anti-fluorescein scFv for test phase. | |||
*Ultimately use anti-rabbit scFv for system. | |||
==Device Depiction== | ==Device Depiction== | ||
<gallery> | <gallery> | ||
| Line 13: | Line 21: | ||
==Current Status== | ==Current Status== | ||
[[../Experiments#Jenny/|Experiments Status]] | |||
<b>FWD for Testing scFvs using anti-FLAG</b> | |||
CATCAT GAATTC GCGGCCGC T TCTAGA TG GACGTCGTTATGACTCAAACACC | |||
-rand- -EcoRI --NotI-- -XbaI[---] Primer | |||
start | |||
*Primers have been designed for biobrick format. | *Primers have been designed for biobrick format. | ||
*pCT-4-4-20, pCT302 S101A, pCT-4M5.3 anti-fluorescein plasmids miniprepped | *pCT-4-4-20, pCT302 S101A, pCT-4M5.3 anti-fluorescein plasmids miniprepped | ||
| Line 23: | Line 39: | ||
==Experiments== | ==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/]] | |||
==Open Issues/Questions== | ==Open Issues/Questions== | ||
Latest revision as of 22:33, 14 August 2005
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
Device Parts
Current Status
[[../Experiments#Jenny/|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/]]
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/]]
