Team:Warsaw/Project/secretion

Theoretical basis
Free living bacteria make use of diverse transport and secretion systems. In gram negative bacteria type I secretion systems are very common e.g. the well characterized hemolysin secretion system. hemolysin is responsible for pathogenicity of some E. coli strains. Obtaining a fusion of protein of interest with hemolysin C-terminus results in secretion of the protein into the growth medium[6] The same method can be used to secret bacterial proteins into the cytoplasm. This has already found its application in the new generation of vaccines[5].

Secretion of p53 into the cytoplasm
To efficiently transport p53 to the eukaryotic cells the endogenic E. coli secretion system responsible for secretion of hemolysin by uropathogenic strains will be used. It is composed of three transport proteins HlyB, HlyD and TolC [10]. TolC is present in the genome of the lab strain derived from K12 strain. To ensure efficient secretion hlyB and hlyD have to be introduced into the genome and proapoptotic protein has to be fused with 60 amino acid long C-terminal sequence of hemolysin. [5].