Potential for adhesion
Patricia Illing
to find something potentially more stable than the YFP.
There are some patogenic strains of E.coli (EHEC and EPEC) that bind
to cell of the gut by inserting their own receptor into the membrane
of the target cell. This receptor is knwon as Tir and the of the
E.coli membrane with which it interacts is known as intimin.
This gives rise to the idea of expressing intimin on the E.coli and
creating a fusion protein of the extracellular tir domain fused to an
E.coli transmembrane protein so as to express it on the E.coli
simultaneously. This would enable our cells to bind one another
(hopefully)
There seems to be a fair bit of research into these proteins and there
are plasmids encoding Intimin which we could hopefully get our hands
on as well as those coding the Intimin binding domain of tir.
a couple of articles worth looking at are:
1. Kenny, B. Enteropathogenic E. coli (EPEC) Transfers Its Receptor
for Intimate Adherence into Mammalian Cells. (1997)
2.Luo, Y. Crystal Structure of enteropathogenic E. coli Intimin-
receptor complex. (2000)
I've uploaded them.
Any thoughts would be very welcome - especially on the fact that this
invovles the creation of our own fusion protein.
Pat
Lei Xing
1. Is intimin expressed on the E.coli membrain as well?
2. How stable is it? how strong the interaction is?
3. I would suggest we use the existing one Fos and Jun bZip, coz we know it is working, we just need to take off the YFP half part from each of the Fos and Jun, which might give better performance to the interaction than what MaGrill did last year. Plus considering the risks we have already had on the floating and light switching system, we would recommend now we use what they have in biobrick library. plus we only have few days for the proposal.
Wat do u guys think?
Cheers!
Lei Xing
Patricia Illing
You’re right there are just too many questions too. Especially when we have such limited time.
I’ve been looking at the biobricks created by McGill and the YFP domains are between the Fos/Jun and the IgA domain. It might be better to try to obtain the plasmids from the Spanish group that don’t contain the YFP – rather than spend time trying to remove it.
Pat
Lei Xing
Patricia Illing
The biobricks for Jun and Fos are BBa_J40004 and BBa_J40006 and contain the YFP domains. I will therefore contact the Spanish group regarding the initial plasmids without the YFP. As they supplied the McGill team with the plasmids last year – hopefully we will be able to obtain them too
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