From OpenWetWare

2-4pm Meeting with Prof. Freemont and Dr Mann

Validation of & discussion about pending test constructs

  • Part J37016 (predator cell test construct):
    • Modelling: Justification of Michaelis-Menten Kinetics
    • Find binding constant for binding curve
  • Part J37022 (aiiA test construct):
    • Do the tags have any influence on activity of enzyme?
    • Look up 3D structure of enzyme (protein databank)
      • Find out about active sites etc. in order to decide on where to place the degradation tag
    • Regarding the half-life of aiiA
      • Better to test both ways with inserting IPTG and after washing out IPTG
  • Part J37015 (prey cell test construct):
    • CRE Recominases
      • Cleavage of DNA -> New joint DNA -> Transcription of cleaved DNA (difficult to achieve in eukaryotes but possible in prokaryotes)
  • Biosensor:
    • AHL with radioactive label ?

Update on Meeting

The 3D structure of AiiA would suggest that it will be possible to insert on either end of the enzyme without hidering the active site. This paper by Kim et al. shows graphical models of AiiA which suggest this. Johnsy