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Using Purified Tyrosine Site-Specific Recombinases In Vitro to Rapidly Construct and Diversify Metabolic Pathways

Lookup NU author(s): Dr Jon Marles-Wright

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Abstract

The site-specific recombinase Cre was previously reported to have in vitro activity. Here, we describe the method of purifying two new tyrosine site-specific recombinases VCre and Dre along with Cre by nickel affinity chromatography. We proved the in vitro function of the VCre and Dre on their respective conditional recombination sites. We also developed a methodology to one-step construct and optimize the productivity of a biosynthetic pathway through the combinatorial integration of promoters into a plasmid-encoded pathway by simply incubating a DNA mixture with recombinase system at 37 °C in vitro.


Publication metadata

Author(s): Liu W, Tuck L, Marles-Wright J, Cai Y

Editor(s): Eroshenko N

Series Editor(s): John M. Walker

Publication type: Book Chapter

Publication status: Published

Book Title: Methods in Molecular Biology: Site Specific Recombinases

Year: 2017

Volume: 1642

Pages: 285-302

Online publication date: 17/08/2017

Acceptance date: 01/01/1900

Series Title: Methods in Molecular Biology

Publisher: Humana Press

Place Published: New York

URL: https://doi.org/10.1007/978-1-4939-7169-5_18

DOI: 10.1007/978-1-4939-7169-5_18

PubMed id: 28815507

Library holdings: Search Newcastle University Library for this item

ISBN: 9781493971695


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