Team:ShanghaiTech/Part Collection
Collection
We mutated several host RBS (ribosome binding site) and host 16s rRNA to get four orthogonal ribosome systems based on the sequences we got. By combining these orthogonal RBS and orthogonal 16s rRNA, we tested their orthogonality to the host ribosomes and to each other. The results showed significant decrease in the interference of exogenous target gene, which is GFP in this case, and host genes, proving the function of the orthogonal ribosome. Since we would like to introduce the orthogonal ribosome to the iGEM community, we develop a toolkit plasmid that contains the report gene GFP under the orthogonal RBS. The superiority of it is that we have also added restriction enzyme cutting sites before and after the GFP sequence. Hence other users can exchange GFP to their target gene easily and select the right clone by simply detecting the fluorescence intensity of the cells.
