Efficient markerless gene deletions in Pseudomonas protegens Pf-5 using a upp-based counterselective system
Xing-Lian Wang1, Si-Yu Dai1, Qing-Jiao Wang1
1School of Pharmacy and Life Sciences, Jiujiang University, Jiujiang, 332000, People's Republic of China.
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.
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