diff --git a/.gitattributes b/.gitattributes index 51fdd51ab1ded0a8a36f82cc04cc674de4ad1dfc..ed16a268209a882558000d5bd7f42c26bbbe7d0a 100644 --- a/.gitattributes +++ b/.gitattributes @@ -6,3 +6,4 @@ assays/Screen[[:space:]]for[[:space:]]antagonistic[[:space:]]interbacterial[[:sp assays/BGC[[:space:]]prediction[[:space:]]using[[:space:]]antiSMASH/dataset/Table[[:space:]]S2.xlsx filter=lfs diff=lfs merge=lfs -text assays/Mini-Tn5TransposonMutantInR401andR569/dataset/.gitkeep filter=lfs diff=lfs merge=lfs -text assays/Mini-Tn5ScreenLossOfR401sGrowthInhibitionRsGMI1600/dataset/.gitkeep filter=lfs diff=lfs merge=lfs -text +assays/Mini-Tn5ForPyoverdineLackofR569/dataset/.gitkeep filter=lfs diff=lfs merge=lfs -text diff --git a/assays/Mini-Tn5ForPyoverdineLackofR569/README.md b/assays/Mini-Tn5ForPyoverdineLackofR569/README.md new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/Mini-Tn5ForPyoverdineLackofR569/dataset/.gitkeep b/assays/Mini-Tn5ForPyoverdineLackofR569/dataset/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/Mini-Tn5ForPyoverdineLackofR569/isa.assay.xlsx b/assays/Mini-Tn5ForPyoverdineLackofR569/isa.assay.xlsx new file mode 100644 index 0000000000000000000000000000000000000000..7d12f4eeed689a3daf17236b1ca1b31321848bc0 Binary files /dev/null and b/assays/Mini-Tn5ForPyoverdineLackofR569/isa.assay.xlsx differ diff --git a/assays/Mini-Tn5ForPyoverdineLackofR569/protocols/.gitkeep b/assays/Mini-Tn5ForPyoverdineLackofR569/protocols/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/Mini-Tn5ForPyoverdineLackofR569/protocols/Mini-Tn5TransposonScreenPyoverdineLackofR569.md b/assays/Mini-Tn5ForPyoverdineLackofR569/protocols/Mini-Tn5TransposonScreenPyoverdineLackofR569.md new file mode 100644 index 0000000000000000000000000000000000000000..9aba88cb408fdf2f8e1a03340e1b946a33b62188 --- /dev/null +++ b/assays/Mini-Tn5ForPyoverdineLackofR569/protocols/Mini-Tn5TransposonScreenPyoverdineLackofR569.md @@ -0,0 +1,3 @@ +## Mini-Tn5 transposon mutant screen for lack of pyoverdine fluorescence of R569 + +R569 mini-Tn5 mutants were cultured in 96 well plates at 25 °C and 180 rpm for five days in 50% TSB medium. For the initial mutant screen, fluorescence was acquired at ðœ†~excitation~=395 nm and ðœ†~emission~=470 nm in a microplate reader (Infinite M200 PRO, Tecan). Out of ~2,000 mutants analysed, the fluorescence-based screening identified twelve R569 mutants that showed severely reduced pyoverdine-specific fluorescence but retained median-like growth behaviour (Thresholds: -6x MAD fluorescence, >-1x MAD Abs~600~). For validation, cultures were pre-grown for five days in 50% TSB before sub-culturing into fresh siderophore medium (see **Table S5**) and growth for five additional days. Finally, bacterial culture density (Abs~600~) was determined, and pyoverdine-specific fluorescence of bacterial culture supernatants was captured at ðœ†~excitation~=410 nm and ðœ†~emission~=500 nm. For comparison between genotypes, fluorescence capacity was calculated by dividing the pyoverdine-specific fluorescence by culture density. \ No newline at end of file diff --git a/assays/Mini-Tn5ScreenLossOfR401sGrowthInhibitionRsGMI1600/isa.assay.xlsx b/assays/Mini-Tn5ScreenLossOfR401sGrowthInhibitionRsGMI1600/isa.assay.xlsx index 5a8d5b32ec6e5e437c862b391b832e4d97436c3b..917c8436bc6717c1ecbf1b59232d204bad7bfb32 100644 Binary files a/assays/Mini-Tn5ScreenLossOfR401sGrowthInhibitionRsGMI1600/isa.assay.xlsx and b/assays/Mini-Tn5ScreenLossOfR401sGrowthInhibitionRsGMI1600/isa.assay.xlsx differ