Protocol for detecting Yersinia pseudotuberculosis nitric oxide exposure during in vitro growth

Rezia Era D Braza; Katherine L Cotten; Kimberly M Davis

doi:10.1016/j.xpro.2022.101760

Protocol for detecting Yersinia pseudotuberculosis nitric oxide exposure during in vitro growth

STAR Protoc. 2022 Dec 16;3(4):101760. doi: 10.1016/j.xpro.2022.101760. Epub 2022 Oct 9.

Authors

Rezia Era D Braza¹, Katherine L Cotten², Kimberly M Davis³

Affiliations

¹ W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA. Electronic address: rbraza1@jhu.edu.
² W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA.
³ W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA. Electronic address: kdavi140@jhu.edu.

Abstract

Yersinia pseudotuberculosis (Yptb) is a bacterial pathogen that causes foodborne illness. Defense against the host antimicrobial gas, nitric oxide (NO), by the bacterial NO-detoxifying gene, hmp, promotes Yptb replication in mouse models of infection. Here, we detail the use of fluorescent signals as readouts for NO exposure within individual cells and subsequent detection of heterogeneity within a population, using single-cell imaging and analysis. This protocol quantifies NO exposure in culture, without capturing the full complexity of the host environment. For complete details on the use and execution of this protocol, please refer to Patel et al. (2021).

Keywords: Cell culture; Microbiology; Microscopy.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Mice
Nitric Oxide
Yersinia pseudotuberculosis Infections* / genetics
Yersinia pseudotuberculosis Infections* / microbiology
Yersinia pseudotuberculosis* / genetics

Substances

Nitric Oxide

Abstract

Publication types

MeSH terms

Substances

Grants and funding