ArticlePloS one2011
Improved expression systems for regulated expression in Salmonella infecting eukaryotic cells.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 23 citations in OpenAlex.
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- Multitask Approach to Localize Rhizobial Type Three Secretion System Effector Proteins Inside Eukaryotic Cells.Plants (Basel, Switzerland) · 2023Article
- Recent advances in bacterial therapeutics based on sense and response.Acta pharmaceutica Sinica. B · 2023Review
- Multilayered safety framework for living diagnostics in the colon.Frontiers in systems biology · 2023Article
- Bacteria-cancer interactions: bacteria-based cancer therapy.Experimental & molecular medicine · 2019Review
- Plant-made Salmonella bacteriocins salmocins for control of Salmonella pathovars.Scientific reports · 2018Article
- The XylS/Pm regulator/promoter system and its use in fundamental studies of bacterial gene expression, recombinant protein production and metabolic engineering.Microbial biotechnology · 2017Review
- Recombinant Salmonella expressing SspH2-EscI fusion protein limits its colonization in mice.BMC immunology · 2017Article
- Article
- Engineering Salmonella as intracellular factory for effective killing of tumour cells.Scientific reports · 2016Article
- Benzoic Acid-Inducible Gene Expression in Mycobacteria.PloS one · 2015Article
- Improved cytotoxic effects of Salmonella-producing cytosine deaminase in tumour cells.Microbial biotechnology · 2015Article
- Synthesis of metallo-β-lactamase VIM-2 is associated with a fitness reduction in Salmonella enterica Serovar Typhimurium.Antimicrobial agents and chemotherapy · 2014Article
- Development of a Fur-dependent and tightly regulated expression system in Escherichia coli for toxic protein synthesis.BMC biotechnology · 2013Article
- Novel tools to analyze the function of Salmonella effectors show that SvpB ectopic expression induces cell cycle arrest in tumor cells.PloS one · 2013Article
- Heterologous viral expression systems in fosmid vectors increase the functional analysis potential of metagenomic libraries.Scientific reports · 2013Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In this work we describe a series of improvements to the Salmonella-based salicylate-inducible cascade expression system comprised of a plasmid-borne expression module, where target gene expression is driven by the P(m) promoter governed by the XylS2 regulator, and a genome-integrated regulatory module controlled by the nahR/P(sal) system. We have constructed a set of high and low-copy number plasmids bearing modified versions of the expression module with a more versatile multiple cloning site and different combinations of the following elements: (i) the nasF transcriptional attenuator, which reduces basal expression levels, (ii) a strong ribosome binding site, and (iii) the Type III Secretion System (TTSS) signal peptide from the effector protein SspH2 to deliver proteins directly to the eukaryotic cytosol following bacterial infection of animal cells. We show that different expression module versions can be used to direct a broad range of protein production levels. Furthermore, we demonstrate that the efficient reduction of basal expression by the nasF attenuator allows the cloning of genes encoding highly cytotoxic proteins such as colicin E3 even in the absence of its immunity protein. Additionally, we show that the Salmonella TTSS is able to translocate most of the protein produced by this regulatory cascade to the cytoplasm of infected HeLa cells. Our results indicate that these vectors represent useful tools for the regulated overproduction of heterologous proteins in bacterial culture or in animal cells, for the cloning and expression of genes encoding toxic proteins and for pathogenesis studies.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.