R401pvdY Complementation assay and protocol

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 assays/Mini-Tn5ScreenLossOfR401sGrowthInhibitionRsGMI1600/dataset/.gitkeep filter=lfs diff=lfs merge=lfs -text
 assays/Mini-Tn5ForPyoverdineLackofR569/dataset/.gitkeep filter=lfs diff=lfs merge=lfs -text
 assays/Mini-Tn5IdentificationofIntegrationSitesR401andR569/dataset/.gitkeep filter=lfs diff=lfs merge=lfs -text
+assays/ComplementationR401pvdY/dataset/.gitkeep filter=lfs diff=lfs merge=lfs -text

assays/ComplementationR401pvdY/protocols/ComplementationR401∆pvdy.md

+## Complementation of R401 ∆pvdy
+
+Complementation of R401 *∆pvdy* was conducted by expressing the coding region of R401 *pvdY* under its putative native promoter from a low-copy plasmid in the *∆pvdy* mutant background. Plasmid construction was conducted using Gibson assembly as described before. In brief, the coding region of R401 *pvdY* was co amplified with a 1.5 kb region upstream of the gene and ligated into a linearized pSEVA22l (18) vector backbone. This vector was integrated into *E. coli* BW29427 cells and subsequently integrated into R401 *∆pvdy via* bi-parental conjugation.
+
+(18) E. Martínez-García, *et al.*, SEVA 3.0: an update of the Standard European Vector Architecture for enabling portability of genetic constructs among diverse bacterial hosts. *Nucleic Acids Res* **48**, D1164–D1170 (2020).
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