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Multiprotein complex production in E. coli: the SecYEG-SecDFYajC-YidC Holotranslocon

Research output: Chapter in Book/Report/Conference proceedingChapter in a book

Original languageEnglish
Title of host publicationHeterologous Gene Expression in E.coli
Subtitle of host publicationMethods and Protocols
EditorsNichola A Burgess-Brown
Publisher or commissioning bodySpringer
Pages279-290
Number of pages12
ISBN (Electronic)9781493968879
ISBN (Print)9781493968855
DOIs
DateAccepted/In press - 8 Sep 2016
DateE-pub ahead of print - 4 May 2017
DatePublished (current) - 2017

Publication series

NameMethods in Molecular Biology
PublisherSpringer New York
Volume1586
ISSN (Print)1064-3745

Abstract

A modular approach for balanced overexpression of recombinant multiprotein complexes in E. coli is described, with the prokaryotic protein secretase/insertase complex, the SecYEG-SecDFYajC-YidC holotranslocon (HTL), used as an example. This procedure has been implemented here in the ACEMBL system. The protocol details the design principles of the monocistronic or polycistronic DNA constructs, the expression and purification of functional HTL and its association with translating ribosome nascent chain (RNC) complexes into a RNC-HTL supercomplex.

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  • Full-text PDF (accepted author manuscript)

    Rights statement: This is the author accepted manuscript (AAM). The final published version (version of record) is available online via Springer at https://link.springer.com/protocol/10.1007%2F978-1-4939-6887-9_18. Please refer to any applicable terms of use of the publisher.

    Accepted author manuscript, 601 KB, PDF document

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