Multiple genes deletion based on Cre-loxP marker-less gene deletion system for the strains from the genus of Pectobacterium.
Pectobacterium
Antibiotic marker
Cre-loxP multiple genes deletion
Journal
Biotechnology letters
ISSN: 1573-6776
Titre abrégé: Biotechnol Lett
Pays: Netherlands
ID NLM: 8008051
Informations de publication
Date de publication:
31 Jul 2024
31 Jul 2024
Historique:
received:
18
04
2024
accepted:
07
07
2024
revised:
03
06
2024
medline:
31
7
2024
pubmed:
31
7
2024
entrez:
31
7
2024
Statut:
aheadofprint
Résumé
To introduce the Cre-loxP system for constructing marker-less multiple-gene deletion mutants in Pectobacterium, overcoming limitations of antibiotic markers and enhancing the understanding of pathogenic mechanisms. Firstly, a plasmid named pEX18-Cre, containing a sacB sucrose suicide gene, was constructed to express Cre recombinase in Pectobacterium. Secondly, a mutant in which the loxP-Km fragment replaced the target gene was obtained through homologous recombination double-crossover with the chromosome. Finally, pEX18-Cre was introduced into the mutant to excise the DNA between the loxP sites, thereby removing the markers and achieving multiple gene deletions. By utilizing the Cre-loxP system, we successfully constructed multiple marker-less gene deletion mutants in Pectobacterium strains. The Cre-loxP system efficiently creates marker-less multiple-gene deletion mutants, enhancing the study of Pectobacterium pathogenic mechanisms by overcoming antibiotic marker limitations.
Identifiants
pubmed: 39083114
doi: 10.1007/s10529-024-03518-8
pii: 10.1007/s10529-024-03518-8
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : National Key Research and Development Program of China
ID : 2021YFC2600602
Organisme : National Natural Science Foundation of China
ID : 32302321
Organisme : Key Laboratory in Science and Technology Development Project of Suzhou
ID : SNG2020045
Informations de copyright
© 2024. The Author(s), under exclusive licence to Springer Nature B.V.
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