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pET expression vector customized for efficient seamless cloning

Dobrijevic, D; Nematollahi, LA; Hailes, HC; Ward, JM; (2020) pET expression vector customized for efficient seamless cloning. Biotechniques 10.2144/btn-2020-0101. Green open access

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Abstract

Here we present a modification of the widely used pET29 expression vector for use in rapid and straightforward parallel cloning via a gene replacement and Golden Gate strategy. The modification can be applied to other expression vectors for Gram-negative bacteria. We have used the modified vectors to clone large numbers of bacterial natural enzyme variants from genomic and metagenomic sources for applications in biocatalysis.

Type: Article
Title: pET expression vector customized for efficient seamless cloning
Location: England
Open access status: An open access version is available from UCL Discovery
DOI: 10.2144/btn-2020-0101
Language: English
Additional information: This article is distributed under the terms of the Creative Commons Attribution License 4.0 which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. To view a copy of the license, visit http://creativecommons.org/licenses/by/4.0/.
Keywords: Golden Gate, expression in E. coli, pET vectors, sacB, seamless cloning
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > Dept of Chemistry
URI: https://discovery.ucl.ac.uk/id/eprint/10114537
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