David Mead
Lucigen Corporation, USA
Title: A novel platform for expressing multi-subunit protein complexes and pathways
Biography
Biography: David Mead
Abstract
Circular plasmids frequently are difficult to use for expression of multiple or large genes, repetitive or toxic proteins and pathways or multi-component gene circuits. The inherent supercoiling of circular plasmids imparts instability forrnthese and other complex gene sequences. The pJAZZ plasmid is a unique linear cloning vector that tolerates nearly any DNA sequence including AT-rich genes and highly repetitive sequences that are impossible to capture in typical circular vectors. pJAZZ has a cloning capacity of up to 40 kb and lacks the cloning bias inherent to circular plasmids, enabling the assembly of complex multi-gene systems. We are developing pJAZZ expression vectors for a variety of applications including expression of mega Dalton proteins, expression of multiple proteins (up to six currently) and production of metabolites from prokaryotic pathways. The vector includes improved light-inducible expression cassettes that have distinct advantages over small-moleculerninduction such as: Precise spatial control of expression; instantaneous initiation and termination of inducing agent; tunability and lack of toxicity or cross-reactivity. The light inducible gene circuit has proven to be significantly more efficient in the linear pJAZZ backbone than in circular vectors.
