Construction and characterization of a temperature-sensitive pRC4 replicon for Rhodococcus and Gordonia.
Molecular docking
Molecular dynamics simulation
Non-replicating plasmid
RepB
Shuttle vector
Site directed mutagenesis
Journal
Gene
ISSN: 1879-0038
Titre abrégé: Gene
Pays: Netherlands
ID NLM: 7706761
Informations de publication
Date de publication:
15 Nov 2023
15 Nov 2023
Historique:
received:
17
10
2023
revised:
08
11
2023
accepted:
13
11
2023
pubmed:
18
11
2023
medline:
18
11
2023
entrez:
17
11
2023
Statut:
aheadofprint
Résumé
Temperature-sensitive plasmids are useful for genome engineering and several synthetic biology applications. There are only limited reports on temperature-sensitive plasmids for Rhodococcus and none for Gordonia. Here, we report the construction of a temperature-sensitive pRC4 replicon that is functional in Rhodococcus and Gordonia. The amino acid residues were predicted for the temperature-sensitive phenotype in the pRC4 replicon using in silico methods and molecular simulation of the DNA-binding replication protein with the origin of replication. The amino acid residues were mutated, and the temperature-sensitive phenotype was validated in Gordonia sp. IITR100. Similar results were also observed in Rhodococcus erythropolis, suggesting that the temperature-sensitive phenotype was exhibited across genera.
Identifiants
pubmed: 37977321
pii: S0378-1119(23)00831-4
doi: 10.1016/j.gene.2023.147990
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
147990Informations de copyright
Copyright © 2023 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.