Journal: bioRxiv

Article Title: Salmonella succinate utilisation is inhibited by multiple regulatory systems

doi: 10.1101/2022.12.21.521472

Figure Lengend Snippet: ( A ) the growth of a collection of S . Typhimurium 4/74 mutants lacking regulatory proteins was assessed on solidified M9+Succ agar and in liquid M9+Succ medium (in microplates), revealing the Δ pnp (JH3649). ( B & C ) Chromosomal complementation of the Δ rpoS mutation: a copy of rpoS (including its native promoter, bent arrow), linked to the cat Cm resistance gene was inserted in the non-transcribed pseudogene STM474_1565 of 4/74 WT (strain 4/74 rpoS 2X , SNW226) and of Δ rpoS ( rpoS + rpoS , JH4160). The STM474_1565 gene is also known as STM1553 or SL1344_1483 . RpoS-dependent phenotypes were assessed for each strain: growth was tested in M9+Succ ( B ) and RDAR phenotype was tested on Congo Red agar plates, confirming the Succ - RDAR + of the complemented strain Δ rpoS + rpoS . ( D ) The growth of the novel Succ + mutants identified (presented in ) was tested on solidified M9 minimal medium supplemented with 40 mM citrate, malate or fumarate. The growth was assessed with biological triplicates after the indicated incubation time (37°C) and the growth of each mutant was compared to 4/74 WT (Succ - ) and Δ rpoS (Succ + ).

Article Snippet: The GFP signal was measured with a FLUOStar Omega plate reader (BMG Labtech) with 200 µL of bacterial suspension per well in black microplates (Greiner #655090).

Techniques: Mutagenesis, Incubation