Team:HokkaidoU Japan/Test

Dr. E.coli : The smallest protein injector in the world

There are several transfection methods to introduce nucleic acid into cells, however methods to introduce proteins directly into cells are complicated.

Unlike introducing DNA, which changes cell genetic makeup permenantly, proteins can act at pinpoint acuracy. And afterwards be degraded leaving no permanent information in the target cell,

Our mission was to find an easy way to introduce desired proteins in the cell. Speficaly into eucaryotic cell. In order to do this we chose Type 3 Secreation System. Find in Salmonella and EPEc. These bacteria are pathogenic and working with them is problematic. To make things easier and safer we ordered DNA fragment which contains T3SS`s syringe part. And introduce it into E.coli.

We also needed protein to secreat. We desined it by ataching secreation signal to desired protein. Our choice was GFP. Also added NLS to make it go to nucleus.

Currently we will test tjis system on liver cancer cells to see if GFP will localize into nucleus.