Team:Warsaw/Project/cytoplasm

Secretion of proteins into the cytoplasm

Theoretical basis

Bacterial systems of protein secretion

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].

Details

The cytoplasmic operon.

Fig 4. Overview of endosomal detection operon. It is composed of YFP – Yellow Fluorescent Protein, tetR – repressor of a tetracycline resistance operon, cro box regulatory sequence from phage and λ desired genes and genes specific for desired function of bacterial strain. In this case p53 – apoptosis inducer.

Cytoplasmic operon, like the invasion operon, is a bistable switch. The regulatory elements are: synthetic promoter inhibited by TetR, promoter inhibited by araC, araC protein (inactivated by L-arabinose) and TetR protein (repressor of tetracycline resistance gene inactivated by tetracycline). [18] This switch controls genes expressed in the cytoplasm. Cro box confines the gene expression to the cytoplasm only. Cro protein bound to it stops the transcription while the endosome detection operon is expressed. The yellow florescence protein is expressed to allow detection of the cytoplasmic operon activity. The set of genes in the cytoplasmic operon depends on the desired functionality of bacterial strain. In case of antitumor therapy could be the p53 protein with mitochondrial leader sequence. In the strain capable of conjugation with mitochondria cytoplasmic operon will control genes responsible for the conjugation process.