IGEM:IMPERIAL/2007/Calendar/2007-7-30

We are in Week 4 of the iGEM project.

name=IGEM:IMPERIAL/2007/Calendar date = 2007/08/01 view=threemonths format=%name/%year-%month-%day weekstart=7

Debriefing
The debrief session was attended by Professor Freemont. A number of issues were raised; these are described below.

Projects
The work done on the various projects was reported at the debriefing session.

Biofilm

 * General protocols available for E Coli experimentation were recorded.
 * Work was started on protocols specific to the Biofilm project
 * It was reported that 4 constructs would be employed. Also, it was suggested, that due to some parallels with last year's Imperial project, these established constructs could already be exploited.
 * More work needs to be done in terms of reporter mechanisms, e.g. DSRed.
 * This project proceeds by employing "test" constructs until the required HRP constructs are made available.

Problems/considerations
 * Can E Coli Polymerase be employed in the in-vitro/in-veso system, or is T7 the only possibility?
 * DSRed, was suggested as a possible reporter system; especially since this would already be characterized from one of the other projects. Are there other reasons which could/could not warrant its use in the Biofilm project? - especially, consider time-scale of operation.

HRP

 * The constructs to be pursued further were discussed.
 * Relevant protocols to be developed per device.

In-Veso

 * Email Christina from the "Vesicles" Division concerning considerations/issues/problems raised including:
 * Stability/Rigidity of vesicles
 * No. of ribosomes possibly accommodated by vesicles - in other words, size considerations.
 * Polymerase restrictions - only T7 employable?
 * Methods of fabrication
 * Materials employed - particular w.r.t. phospholipid compostion/mixture used.
 * Storage of vesicles, especially at temperatures of 4 degrees Celsius.
 * Communication methods across membrane.
 * Alternative means of forming pores within membrane, since a-hemolysin toxin is not permitted during our project work.
 * Also, included in this email, should be the relevant research papers on which our methods/project work to date has been based.

Cell-by-Date

 * Proposed new reporter mechanism, involving DSRed - inherent long degradation time.
 * Issues overlapping with In-Vitro/In-Veso systems, e.g. non-T7 polymerase, etc.
 * Cmv was initially proposed as a constitutive promoter; however, this can only be used successfully within mammalian cells; not prokaryotic cells.
 * The list of experiments to be conducted should be completed.
 * (other issues covered? input required)

Ambitions by end of week

 * Start production of media within lab.
 * Hopefully start cloning for E Coli System.
 * Organize division of labour between different projects, and, within the projects themselves, especially w.r.t labwork.