IGEM:IMPERIAL/2009

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** Restriction sites be inserted by codon changes; GeneArt optimise constructs to remove restriction sites all the time, so putting them in should be OK
** Restriction sites be inserted by codon changes; GeneArt optimise constructs to remove restriction sites all the time, so putting them in should be OK
** Is not native; if it's expressed somehow by the cell, it'll suicide on its own
** Is not native; if it's expressed somehow by the cell, it'll suicide on its own
-
Saint says:
 
# Regulation of expression:
# Regulation of expression:
** Needs to be near 100% off when off; even a low level of expression will destroy your cells long before any product is produced
** Needs to be near 100% off when off; even a low level of expression will destroy your cells long before any product is produced

Revision as of 11:44, 23 July 2009

iGEM 2009 - Imperial College London Team

Work in progress ...

Contents

ToDo & Deadlines

Useful Links


Team Roles

Change your role when applicable

Charles - Cell Death
Dave - Cell Death/ Applications
Dineka - Encapsulation - biosynthetic pathway of alginate
James - Encapsulation - biosynthetic pathway of alginate
Kun - Encapsulation - biosynthetic pathway of alginate
Nuri - Cell Death
Royah - Compounds/BioBricks (Killing Strategy)/Timers (if time)/General
Tianyi - Compounds/Applications

Wiki Updates


Synthetic Biology @ Imperial

iGEM resources

Advisor Contributions

Schumann lab from Uni. Beyreuth, DE have done some interesting work on using spores to direct antigens to the gut - to act as vaccines. Sporulation guys might also be interested in this paper, describing the B. sub coat protein (and how it's hilariously complex but not all required). Oh, and subtilis spores will germinate in the gut (probably), justifying using the killswitch!

Killswitch guys, I think perhaps looking into recombinases as opposed to restriction enzymes would be useful as they won't act on host DNA. Xer and Dif sites will recombine with themselves in presence of the required enzyme, excising any genes between them - you could flank genes with them, then express the enzyme to chop your construct up. Sites are required to be within ≈5kb of each other, I think, so random ones on host DNA shouldn't be affected. It might take a while to work so look into the time; could be useful as a fallback, anyway.

Biobrick images you can use if you need (advisors like named/labelled circuit diagrams!): Image:P.png Image:R.png Image:G.png Image:T.png

~ Tom Adie 15:44, 20 July 2009 (EDT)

Killing guys, things to keep in mind for the restriction enzymes...

  1. Restriction enzyme:
    • Cuts at short sequences; makes it easier to insert (see next point) and will cut in the genome and plasmid more often by chance
    • Restriction sites be inserted by codon changes; GeneArt optimise constructs to remove restriction sites all the time, so putting them in should be OK
    • Is not native; if it's expressed somehow by the cell, it'll suicide on its own
  1. Regulation of expression:
    • Needs to be near 100% off when off; even a low level of expression will destroy your cells long before any product is produced
    • You may want to look for a less-efficient enzyme so small leakage wouldn't be that bad; or an enzyme that is degraded relatively quickly
    • Bistable switch with no leakage might be good; or flippase regulation! My pet idea for the bioremediation project last year =P

You can also insert sites after the transcription terminators in all your genes to ensure full destruction of the construct, and maybe add your sites into normal bricks (terminators etc.) and put them up as variants.

~ Tom Adie 11:41, 23 July 2009 (EDT)


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