Efficient markerless gene deletions in Pseudomonas protegens Pf-5 using a upp-based counterselective system
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.
KeywordsCounterselective system Markerless gene deletion Pseudomonas protegens Upp
This work was supported by the National Natural Science Foundation of P. R. China (31760534) to Dai-ming Zha.
Compliance with ethical standards
Conflict of interest
All authors declare no conflict of interest.
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