1. Efficient markerless gene deletions in Pseudomonas protegens Pf-5 using a upp-based counterselective system.
- Author
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Wang, Xing-lian, Dai, Si-yu, Wang, Qing-jiao, Xu, Hui-nan, Shi, Hong-qiu, Kang, Yu-bin, and Zha, Dai-ming
- Subjects
DELETION mutation ,PSEUDOMONAS ,PLASMIDS - Abstract
Objectives: Developing a counterselective system for efficient markerless gene deletions in biocontrol strain P. protegens Pf-5. Results: 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. Conclusion: 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. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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