Vibrio natriegens with Xtreme Protein Expression and Secretion System Optimization
After years of domination, E. coli has finally faced a challenge with the emergence of Vibrio natriegens, a gram-negative marine bacteria that possesses the potential to become the next generation chassis organism for synthetic biology as it can divide at its best at seven minute intervals.
Inspired by the possibilities that V. natriegens offers, our team Aalto-Helsinki is determined to take on the challenge of improving the bacterium in protein expression and secretion, utilizing the putative signal sequence that V. natriegens has while also producing bacteriophage tools for the strain. We aim to produce single-chain variable fragments (scFv) as a proof-of-concept that V. natriegens can be utilized as a productive cell factory. Our motivation behind our project is to advance synthetic biology and the production of proteins for the benefit of science & iGEM community, and for the society at large.
Our project aims to research and improve the protein expression of Vibrio natriegens, a gram-negative marine bacteria which has an immense potential to be the next-generation workhorse for biotechnology and synthetic biology due to its incredibly fast dividing rate (seven minutes at best). We strive to develop V. natriegens as a cell factory in versatile protein production, and to enhance the bacterium’s secretion pathways to secrete proteins directly into the medium, especially via the twin-arginine translocation (tat) pathway.
To demonstrate the potential of V. natriegens as a secretive cell factory, we plan to express single-chain variable fragments (scFv) of antigens or antibodies. ScFvs are commonly used in flow cytometry, immunohistochemistry, and as antigen-binding domains of artificial T-cell receptors. ScFvs have also great potential in constructing immunotoxins, therapeutic gene delivery, and anticancer intrabodies for therapeutic purposes.
We also plan to prove an identified putative secretion signal that V. natriegens possesses, and to define the secretion pathway it utilises, in addition to researching how it affects the protein secretion.
In addition, we plan to try out infecting V. natriegens with bacteriophages, to see if phages could serve as a possible integration, lysis and mutation tool in the future with V. natriegens.