An expanded library of orthogonal split inteins enables modular multi-peptide assemblies.
Biotechnology
/ methods
Cloning, Molecular
Feasibility Studies
Fluorescence
Gene Library
Genes, Reporter
/ genetics
Inteins
/ genetics
Luminescent Proteins
/ chemistry
Peptides
Plasmids
/ genetics
Protein Engineering
/ methods
Protein Splicing
Recombinant Fusion Proteins
/ chemistry
Transcription Factors
/ genetics
Transformation, Bacterial
Red Fluorescent Protein
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
23 03 2020
23 03 2020
Historique:
received:
04
11
2019
accepted:
26
02
2020
entrez:
7
4
2020
pubmed:
7
4
2020
medline:
22
7
2020
Statut:
epublish
Résumé
Inteins are protein segments capable of joining adjacent residues via a peptide bond. In this process known as protein splicing, the intein itself is not present in the final sequence, thus achieving scarless peptide ligation. Here, we assess the splicing activity of 34 inteins (both uncharacterized and known) using a rapid split fluorescent reporter characterization platform, and establish a library of 15 mutually orthogonal split inteins for in vivo applications, 10 of which can be simultaneously used in vitro. We show that orthogonal split inteins can be coupled to multiple split transcription factors to implement complex logic circuits in living organisms, and that they can also be used for the in vitro seamless assembly of large repetitive proteins with biotechnological relevance. Our work demonstrates the versatility and vast potential of an expanded library of orthogonal split inteins for their use in the fields of synthetic biology and protein engineering.
Identifiants
pubmed: 32251274
doi: 10.1038/s41467-020-15272-2
pii: 10.1038/s41467-020-15272-2
pmc: PMC7090010
doi:
Substances chimiques
Luminescent Proteins
0
Peptides
0
Recombinant Fusion Proteins
0
Transcription Factors
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1529Subventions
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/N007212/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/S018875/1
Pays : United Kingdom
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