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

Wang, Xinglian; Dai, Siyu; Wang, Qing-Jiao; Xu, Hui-Nan; Shi, Hong-Qiu; Kang, Yubin; Zha, Daiming

doi:10.1007/s10529-019-02772-5

Cited by 3 publications

(2 citation statements)

References 25 publications

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“…The 1496 bp upstream homologous arm pueAU (bp À1500 to À5 relative to the translational start site) and the 1544 bp downstream homologous arm pueAD (bp +1844 to +3387 relative to the translational start site) of pueA were, respectively, amplified by PCR using primers pueAU-U-BamHI/pueAU-L-HindIII and pueAD-U-HindIII/pueAD-L-XbaI (Table S2) from the genomic DNA of Pf-5. Then, pueAU and pueAD were cloned into the suicide plasmid pJQ200UPP (Wang et al 2020) to obtain the recombinant plasmid pJQΔpueA (Table S1).…”

Section: Construction Of Plasmidsmentioning

confidence: 99%

“…Four markerless deletion mutants of pueA, pueB and apr-DEF, that is, Pf3202 (DpueA derivative of Pf5139), Pf3205 (DpueB derivative of Pf5139), Pf3202+05 (DpueAB derivative of Pf5139) and Pf3206-08 (DaprDEF derivative of Pf5139) (Table S1) were constructed through a upp-based counterselective system using Pf5139 (Dupp derivative of Pf-5) as their parental strain based on our former study (Wang et al 2020).…”

Section: Construction Of Strainsmentioning

confidence: 99%

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Two polyurethanases PueA and PueB are major extracellular lipases partly secreted by the mediation of their cognate ABC exporter AprDEF in Pseudomonas protegens Pf‐5

Zha

Wang

Xiao

et al. 2021

Lett Appl Microbiol

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Significance and Impact of the Study: We investigated the additional function of two polyurethanases PueA and PueB from Pseudomonas protegens Pf-5, and demonstrated that they are major extracellular lipases that belong to the members of subfamily I.3 lipases, involved in the uptake of oil in Pf-5, of which PueA plays a more important role in extracellular lipase activity. Furthermore, their cognate ABC exporter AprDEF can partly mediate the extracellular secretion of PueA and PueB in Pf-5 and heterologous Escherichia coli BL21(DE3). Lastly, PueA and PueB can also be secreted into the extracellular environment without the help of their cognate AprDEF in Pf-5 and BL21(DE3).

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Section: Construction Of Plasmidsmentioning

confidence: 99%

Section: Construction Of Strainsmentioning

confidence: 99%