4G cloning: rapid gene assembly for expression of multisubunit protein complexes in diverse hosts.

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State: Public
Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_BA8E24BE4E11
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
4G cloning: rapid gene assembly for expression of multisubunit protein complexes in diverse hosts.
Journal
Life science alliance
Author(s)
Taschner M., Dickinson J.B., Roisné-Hamelin F., Gruber S.
ISSN
2575-1077 (Electronic)
ISSN-L
2575-1077
Publication state
Published
Issued date
01/2025
Peer-reviewed
Oui
Volume
8
Number
1
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Abstract
Multisubunit protein complexes are central to many cellular processes, and studying their activities and structures in vitro requires reconstitution via recombinant expression and purification. Obtaining targets at sufficient purity and scale typically involves screening several protein variants and expression hosts. Existing cloning strategies enable co-expression but are often time-consuming, labor-intensive, and host-specific, or involve error-prone steps. We present a novel vector set and assembly strategy to overcome these limitations, enabling expression construct generation for multisubunit complexes in a single step. This modular system can be extended to additional hosts or include new tags. We demonstrate its utility by constructing expression vectors for structural maintenance of chromosomes complexes in various hosts, streamlining workflows, and improving productivity.
Keywords
Cloning, Molecular/methods, Genetic Vectors/genetics, Multiprotein Complexes/genetics, Multiprotein Complexes/metabolism, Recombinant Proteins/genetics, Recombinant Proteins/metabolism, Escherichia coli/genetics, Escherichia coli/metabolism, Protein Subunits/genetics, Protein Subunits/metabolism, Humans, Gene Expression/genetics
Pubmed
Web of science
Open Access
Yes
Create date
09/12/2024 15:10
Last modification date
21/01/2025 7:26
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