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URL: https://dx.doi.org/10.17504/protocols.io.bdmqi45w

Authors: Kristoffer Bach Falkenberg, Cristina Hernandez Rollan, Maja Rennig, Andreas Birk Bertelsen, Morten Norholm

Summary: Lytic polysaccharide monooxygenses (LPMOs) are enzymes that play a critical role in breaking the chemical bonds of the most abundant polymers found in recalcitrant biomass, such as cellulose and chitin. LyGo cloning (Lytic Polysaccharide Monooxygenase Golden Gate cloning) is a versatile heterologous expression platform for LPMOs, which is compatible with cloning both PCR products and synthetic gene fragments with a simple 15-minute assembly step. The method allows for parallel construction of multiple expression vectors, enabling exploration of several expression strategies. The open-source LyGo collection consists of vectors for some of the most relevant model organisms used for protein production in both academic and industrial settings.This protocol describes how to clone LyGo fragments into LyGo vectors.

Affiliations: Technical University of Denmark, Technical University of Denmark, Technical University of Denmark, Technical University of Denmark, Technical University of Denmark

Version: 1

Publication Date: 2020

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