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Efficient markerless gene deletions in Pseudomonas protegens Pf-5 using a upp-based counterselective system.

Authors
  • Wang, Xing-Lian1
  • Dai, Si-Yu1
  • Wang, Qing-Jiao1
  • Xu, Hui-Nan1
  • Shi, Hong-Qiu1
  • Kang, Yu-Bin2
  • Zha, Dai-Ming3
  • 1 School of Pharmacy and Life Sciences, Jiujiang University, Jiujiang, 332000, People's Republic of China. , (China)
  • 2 School of Nursing, Jiujiang University, Jiujiang, 332000, People's Republic of China. , (China)
  • 3 School of Pharmacy and Life Sciences, Jiujiang University, Jiujiang, 332000, People's Republic of China. [email protected] , (China)
Type
Published Article
Journal
Biotechnology Letters
Publisher
Springer-Verlag
Publication Date
Nov 28, 2019
Identifiers
DOI: 10.1007/s10529-019-02772-5
PMID: 31781926
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Developing a counterselective system for efficient markerless gene deletions in biocontrol strain P. protegens Pf-5. We successfully implemented a markerless deletion of upp in Pf-5 to obtain the 5-FU resistant strain Pf5139. With this strain, we performed markerless gene deletions for each component of Gac/Rsm system and a 17 kb DNA fragment with the deletion ratio of 20 to 50%, and efficiently constructed a strain with triple deletions based on the suicide plasmid pJQ200UPP. In addition, there is no obvious connection between the deleted fragment length and the deletion ratio. The upp-based counterselective system in this study is efficient and valuable for markerless gene deletions in Pf-5, indicating that it has great potential in the study of gene function and in the application of genome reduction for Pseudomonas strains.

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