Newcastle University/11 September 2008

Minutes

Minute Taker = Jess Thompson

Time: 16:00 – 17:00

Attended by:

Neil Wipat - supervisor

Jennifer Hallinan - supervisor

Matthew Pocock

Leendert Hamoen - supervisor

Jan-Willem Veening – advisor

Khanh Tang - dry lab student

Ria Chalder – wet lab student

Jess Thompson – wet lab student

Absent:

Megan Aylward

Nina Nielsen-Dzumhur

Morgan Taschuk

Mark Wappett

Colin Harwood

James Lawson

Michael T. Cooling

Meeting Content

Agenda 1: New student

Jen introduces Khanh Tang to the team. She is a student who will be work on the computational side of the project.

Agenda 2: Current state of laboratory work

Jan-Willem, Ria and Jess are pleased to announce that the ncl08 fragment has been successfully transformed stably into B. subtilis and that the bacteria respond to subtilin. Flow cytometry showed a doubling in fluoresces when 1% subtilin supernatent was used, and a further dramatic increase when 10% subtilin supernatent was added. Jen, Anil and Matt are interested in finding out the exact concentration of the subtilin (as it was obtained by the relatively crude method of centrifuging cells and pipetting off the supernatent). Jan-Willem says he will look into this.

Jan-Willem is interested in using quantitative time-lapse as another method for quantifying the response. The images produced from this would be ideal for the team poster in November. Leendert also suggests possibly using microtitre plates to view the GFP fluorescence. Jan-Willem agrees this would be feasible. Matt is interested in this approach as it would be for for modelling purposes.

Anil is interested in the possibility of making the cells express enough fluorescence to be able to see with the naked eye. This would require ligating more reporter genes or more promotors into the correct place in the B. subtilis genome. Jan-Willem agrees that this would be ideal, but expresses concerns over the amount of time available to do this.

Agenda 3: Proteomics

Jan-Willem is expresses interest in conducting proteomics work to determine the physical responses of B. subtilis to subtilin. He mentions this is important as microscopy showed that some of the cells responded well but others did not. This would also allow a direct correlation between the numbers of protein molucules in the medium and the numbers of cells fluorescing to be determined. Lots of sample would be needed to do this.

Jen is interested in getting students back into the laboratory to conduct the proteomics work. This would also possibly allow Morgan to observe the work going on.

Agenda 4: Other future lab work

Jan-Willem would like more microscopy to be performed on the transformed B. subtilis when he returns on 22.09.08. He would also like to test the agr system (when it has been transformed) with Staphylococcus aureus to determine whether it is successfully inducible. This can be done by Ria and Jess whilst he is away.

pJWV021-ncl08 has been checked and is ready to be sent for sequencing. However, pGFPrrnB-ncl08 showed no 2.2kb fragment, so this is to be reisolated and rerestricted to see if it was due to human error and faulty plasmid. When this plasmid and pGFPrrnB-BB107 have been verified as containing their relevant inserts, all three plasmid tubes are to be sent together for sequencing.

Agenda 5: Judging of iGEM teams

Jen asks about current state of the external wiki page. Ria and Jess say that although progress is good, they will need at least one afternoon next week to bring the wet lab section up to date. Jen agrees that this is important as the team is largely judged on its wiki page.

Jen reminds the students that work will need to start soo on the poster and presentation for the Jamboree in November. The talk (of around 5 minutes each) will be done primarily by Ria, Morgan and Jess as other students are currently away.

Jen and Anil mention that a t-shirt logo and banner logo will be needed. Purple is suggested as the colour of choice for the t-shirts. AOB

Jen and Anil will be away on 14th September so will be absent from the 18th September meeting.

Jan-Willem will be away 13th-22nd September and will also be absent for this meeting. It is decided therefore that the meeting will be cancelled for the next week.