Meeting Minutes for July 14, 2006

From 2006.igem.org

July 14th, 2006 Faculty Meeting:

Actions from last week:

1. Megan to look into Avadin

Best way to obtain Avadin, to purchase it, not that expensive, can get it with diff. reporters attached

        Still need source of  Avadin DNA, need DNA sequence 

Will email around the paper, MMS13 is a transmembered protein

2. Jason and Jamie G. found sequence of the vector that guys in Japan will be sending,who have made it so that it can be replicated in e. Coli (6kb total)

        3KB WOULD BE THE CRYPTIC VECTOR THAT THEY ISOLATED FROM THE MAG. BACT. 

        Can cryptic region be isolated from our own mag. bact.? We will look for that?(PLAN B)

        Can incorporation be selected for insertion into a given cell? (Jason will continue to
        look into it)

        End goal would be to make it a part, once we make the 4 cut sites, to make it a std.  
        shuttle vector 

        In final plasmid, incorporated the whole plasmid with the Puc19 

        Idea is to put avadin linked to the C terminus DNA sequence (they put
        luciferase at the C terminus 

a.Look into where we can attach the avadin (we are thinking of using the C terminus)
b.biotin present in the growth media? Could be a problem if we were to tag the mag. bact. with avadin (YES IT IS!!!) Maybe we don’t need it for a growth media
c.Need to reevaluate the magnetotactic bacteria ideas
d.Protein A made by bact. but binds FC regions of proteins
e.Thinks about using invasin!, binds to beta integrin, it would invade and be endocytosed into the cell
f.stimulates uptake by RAC1

3.AHL (Azeem)

       Checked out sigma, didn’t have the sigma acyl group but has other homolysines 

       Might not need to have it, b/c we have the sender and receiver cells almost made

       Contact the igem lab that used AHL 

Update of Sender Cell:

1.Already have one ligation done, have yet to add the promoter
2.Might have to sequence it at this first ligation, before needing to add the promoter
Goal for next week:
1.Have all three put together

Update on AHL sensor:

1.Have to complete ethanol precipitation and ligations

Freeze-Machine:

1.Annie-ligating her two parts
2.Jamie-miniprep that did not have any visible DNA, had to start the miniprep again

Unfreeze:

1.At the step of ligating the parts together

Ways to characterize the parts:

    Ligating GFP to the BBa_P2620