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Team:Cambridge/Quiescence Notes
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* The Lambda phage is the most stable to have an off switch, it does not express itself when needed (?) | * The Lambda phage is the most stable to have an off switch, it does not express itself when needed (?) | ||
* The RNA has a very long half life, it would be hard to strip away the system and put it under a new promotor, but, if possible would be extremely useful | * The RNA has a very long half life, it would be hard to strip away the system and put it under a new promotor, but, if possible would be extremely useful | ||
| + | |||
| + | A new approach to making a quiescence switch would be to control the functionality of the folded RNA instead of its transcription, which proved to be problematic. Some data on the structure of different Rcd can be found here: | ||
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| + | *[http://mic.sgmjournals.org/cgi/reprint/145/8/2135 1999 article by Sharpe et al. Summers being one of the al] | ||
| + | Structure | ||
Revision as of 08:59, 16 July 2010
- [http://mic.sgmjournals.org/cgi/reprint/155/8/2676.pdf The role of FIS in the Rcd checkpoint and stable maintenance of plasmid ColE1] ie good as the toggle control thingy
- [http://www3.interscience.wiley.com/cgi-bin/fulltext/119127730/PDFSTART Timing, self-control and a sense of direction are the secrets of multicopy plasmid stability]
- [http://aem.asm.org/cgi/reprint/65/6/2710.pdf The quiescent-cell expression system for protein synthesis in Escherichia coli]
- [http://apps.isiknowledge.com/full_record.do?product=UA&search_mode=GeneralSearch&qid=12&SID=W11BA3C7bKeHC5PbfoE&page=1&doc=1&colname=WOS ColE1 multimer formation triggers inhibition of Escherichia coli cell division.]
- [http://books.google.com/books?hl=en&lr=&id=OKn60-HZisUC&oi=fnd&pg=PA19&dq=hn-s+nucleoid+bacillus&ots=rstjtFpQ2Z&sig=15uulXPnHz-yyXMvMKRrTXcLk3U#v=onepage&q=h-ns&f=false book about bac genomes]
- [http://www.faqs.org/patents/app/20090004700 D. Summers' Patent Application for Chem. Induction of Quiescence in Bacteria]
Literally no documentation of h-ns and rcd in bacillus.
If we do quiescence, will have to be in e coli with bacillus as a side project. Have to get over IP issues with ucam + e coli. Implementation in ecoli will be fairly easy as it is a working system - all we need to do is brick it, and get it working.
Input from James
- Has to be done in E.Coli, will not work in non Coliform bacteria such as Bacilli.
- Switch on cell division would be very useful, however there are IP issues which must be discussed with David Summers.
- Another constraint is that liquid broth not agar must be used.
- Nobody has looked at it from a microfluidics point of view.
- When done before, the lambda phage takes 3 hours to initiate quiescence after the temperature change.
- The Lambda phage is the most stable to have an off switch, it does not express itself when needed (?)
- The RNA has a very long half life, it would be hard to strip away the system and put it under a new promotor, but, if possible would be extremely useful
A new approach to making a quiescence switch would be to control the functionality of the folded RNA instead of its transcription, which proved to be problematic. Some data on the structure of different Rcd can be found here:
- [http://mic.sgmjournals.org/cgi/reprint/145/8/2135 1999 article by Sharpe et al. Summers being one of the al]
Structure
