Cambridge/Project summary
From 2007.igem.org
Contents |
Introduction
This year's [http://www.ccbi.cam.ac.uk/iGEM2007 Cambridge iGEM team], made up of five physical scientists with a biological bent and six biologists who want to try something a bit different, has been working since the start of July to get to grips with Synthetic Biology, and then to push its frontiers. After extensive brainstorming, we have decided to work on three projects to add to the capabilities of the Registry.
PoPS amplifier
The first strand of our research is the construction of one or more well-characterised PoPS amplifiers, based on a number of protein activators and cognate inducible promoters from a number of E. coli phages investigated by other researchers. At present we are integrating the six promoter sequences and three activator sequences into plasmids; we will then carry out double transformations to generate and characterise the 18 different combinations. [http://www.ccbi.cam.ac.uk/iGEM2007/index.php/Amplifier_project More details on our website...]
B. subtilis as a new chassis
In parallel, we are developing Bacillus subtilis as a Gram-positive chassis for conducting sythetic biology experiments. This involves identifying, obtaining and testing suitable strains, plasmids (especially shuttle vectors between B. subtilis and E. coli), protocols and biobrick parts (promoters, RBSes, reporter proteins and terminators); at present we are testing growth and transformation protocols using different strains and vectors. Gram-positive bacteria have a number of advantages, particularly for protein secretion and chromosomal insertion of DNA. [http://www.ccbi.cam.ac.uk/iGEM2007/index.php/Bacillus_subtilis_SynBio_chassis More details on our website...]
Introducing the Agr signalling system into other organisms
Finally, we are re-engineering the Agr (Accessory Gene Regulator) oligopeptide quorum sensing system from Staphylococcus aureus into a PoPS-compatible intercellular signalling system, with BioBricks for the signal senders and receivers. Initially, we will make use of the B. subtilis chassis to test our system (since S. aureus is also Gram-positive); we also plan to attempt to transfer the system to a Gram-negative chassis (E. coli) using a mutant pore protein to render the outer membrane permeable to oligopeptides and other small molecules. [http://www.ccbi.cam.ac.uk/iGEM2007/index.php/Signalling_project_description More details on our website...]
(last updated 6 August 2007)
[http://www.ccbi.cam.ac.uk/iGEM2007 Our team wiki (external)]
