Salmonella Typhimurium Effector Protein Analysis

Infection experiments were performed using Salmonella enterica serovar Typhimurium (STM) NCTC 12023 strain as WT and isogenic mutant strains (Table 1). Mutagenesis was carried out as described elsewhere^{45 (link)}. In short, strains were constructed using λ Red-mediated mutagenesis and the resistance cassette was removed using FLP-mediated recombination. Mutant strains deficient in effector genes harboring plasmids encoding the corresponding effector fused to HaloTag (Table 2) were used for microscopic analysis.Table 1

Salmonella enterica serovar Typhimurium strains used in this study

Designation	Relevant characteristics	Reference
NCTC 12023	Wild type	Lab stock
MvP388	ΔsscB sseF::FRT	^{52 (link)}
MvP392	ΔsseJ::FRT	^{53 (link)}
MvP503	ΔsifA::FRT	^{54 (link)}
MvP742	ΔsteC::FRT	This study
MvP1900	ΔsseJ::aph ΔsifA::FRT	^{45 (link)}
MvP1944	ΔpipB2::FRT	^{44 (link)}
MvP1948	ΔsseF::aph ΔsifA::FRT	This study
MvP1980	ΔsseF::FRT	^{55 (link)}

Table 2

Plasmids used in this study

Designation	Relevant genotype	Source/reference
p2095	P_sseA::sscB sseF::M45	^{52 (link)}
p2129	P_sseJsseJ::M45	^{41 (link)}
p2621	P_pipB2pipB2::M45	^{18 (link)}
p2643	P_sseA::sscB sseF::HA	^{52 (link)}
p2888	P_sseA::sscB sseF::HA sseG::M45	^{52 (link)}
p3805	P_CMV::LAMP1-miniSOG-mCherry	this study
p3806	P_CMV::LAMP1-tdminiSOG-mCherry	this study
p3991	LAMP1::HaloTag::GFP	^{9 (link)}
p4305	P_sifAsifA::L16::HaloTag::HA	^{16 (link)}
p4118	P_sseAsscB sseF::L16::HaloTag::HA	^{16 (link)}
p4286	P_sseJsseJ::L16::HaloTag::HA	^{16 (link)}
p4295	P_pipB2pipB2::L16::HaloTag::HA	^{16 (link)}
p5059	P_steCsteC::L16::HaloTag::HA	This study
p5065	P_sseA::sscB sseF::3HA	This study
p6172	P_pipB2pipB2 ΔCC::L16::HaloTag::HA	this study
p6173	P_sopD2sopD2 ΔCC::L16::HaloTag::HA	this study
p6187	P_sifAsifA Δ331-336::L16::HaloTag::HA	this study
p6203	P_sseJsseJ ΔCC::L16::HaloTag::HA	this study

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Göser V., Sander N., Schulte M., Scharte F., Franzkoch R., Liss V., Psathaki O.E, & Hensel M. (2023). Single molecule analyses reveal dynamics of Salmonella translocated effector proteins in host cell endomembranes. Nature Communications, 14, 1240.

Publication 2023

Genes Halotag Infection Lamp1 Microscopic Mutagenesis Plasmids Recombination Salmonella enterica Strains

Corresponding Organization :

Other organizations : Osnabrück University

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Variable analysis

independent variables

Salmonella enterica serovar Typhimurium (STM) NCTC 12023 strain as WT and isogenic mutant strains (Table 1)
Mutant strains deficient in effector genes harboring plasmids encoding the corresponding effector fused to HaloTag (Table 2)

dependent variables

Microscopic analysis of the mutant strains

control variables

Wild type Salmonella enterica serovar Typhimurium (STM) NCTC 12023 strain

controls

Positive control: Wild type Salmonella enterica serovar Typhimurium (STM) NCTC 12023 strain
Negative control: Not explicitly mentioned

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