Team:Warsaw/Project

This year's goal of iGEM Warsaw team is to develop 2 foundational techniques for synthetic biologists:

So far all RBS parts was measured with GFP protein.We have measured the RBS parts with various fluorescent proteins and guess what - they are not as standard as we would like them to be. The strength of a RBS part depend on the protein used, because the beginning of the protein influence the mRNA fold. When mRNA is strongly folded ribosome can not start translation. We came up with the idea of RBS parts fused with short 'protein beginnings' - we call this expression adapters.We are working on expression adapters that would provide standardized protein expression from each RBS from community collection. Also we are working on adapters that increase protein expression

Our goal is to make easy and efficient protocol for cell free cloning. It skips most basic step of classical cloning approach - plasmid propagation in bacteria or yeast. This allows cloning of constructs that are toxic to host cells (i.e. nucleases or lysins) and speeds up the whole procedure at least three times. The cell free cloning process involves rolling circle amplification of ligation products by phi29 polymerase. Our protocol is speed-oriented so we'll try to optimize all time consuming steps: DNA digestion, substrate purification, ligation and product amplification. We'll also try to deal with specificity problems of phiX29 polymerase by designing RNA primers optimized for BioBrick amplification.