Evaluation of Gram-negative bacterial infection by a stable and conjugative bioluminescence plasmid in a mouse model

Yao Kuang Huang, Chishih Chu, Chih Hsiung Wu, Chyi Liang Chen, Cheng Hsun Chiu

Research output: Contribution to journalArticle

Abstract

BACKGROUND: The green fluorescence protein (GFP)-associated fluorescence method and the luciferase-associated bioluminescence method are the two major methods for IVIS imaging system to investigate the bacterial infection in animal models. The aim of this study was to evaluate the infection route of Gram-negative bacteria carrying a stable and broad range of conjugative bioluminescence plasmid pSE-Lux1 in a mouse model.

RESULTS: Both encapsulated and non-encapsulated Gram-negative bacteria were used as hosts to evaluate conjugation efficiency and plasmid stability of pSE-Lux1, a recombinant of pSE34 and luxABCDE operon. The plasmid conjugation efficiencies of pSE-Lux1 ranged from 10⁻³ to 10⁻⁷ in various Gram-negative bacteria. Plasmid pSE-Lux1 maintained in Escherichia coli, Klebsiella pneumoniae, and Salmonella enterica serovars Choleraesues (abbreviated S. Choleraesuis) and Typhimurium (S. Typhimurium), than in Acinetobacter baumannii and Serratia marcescens, was shown to be of better stability for at least four days. To investigate systemic bacterial infections, K. pneumoniae strain CG354 was intravenously injected, and then was clearly observed to be non-pathogenic to Balb/c mice for a long-term bioluminescence monitoring for 6 days. For examining dynamic distributions of gastrointestinal tract infection, the invasion protein SipB-deficient mutant OU5045△sipB and OU5046△sipB of S. serovar Typhimurium constructed in this study, compared to wild-type strain OU5045 and its virulence plasmid-less strain OU5046, were of less virulence to mice.

CONCLUSIONS: This is the first study to evaluate the conjugative and stable bioluminescence vehicle system of pSE-Lux1 in a wide range of Gram-negative bacteria, a system that can provide a useful reporter approach to trace systemic and gastrointestinal bacterial infections in a mouse model.

Original languageEnglish
Pages (from-to)78
Number of pages1
JournalJournal of Biomedical Science
Volume21
DOIs
Publication statusPublished - 2014
Externally publishedYes

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Gram-Negative Bacterial Infections
Bioluminescence
Gram-Negative Bacteria
Plasmids
Bacteria
Bacterial Infections
Klebsiella pneumoniae
Virulence
Fluorescence
Acinetobacter baumannii
Serratia marcescens
Salmonella
Salmonella enterica
Operon
Infection
Luciferases
Imaging systems
Escherichia coli
Gastrointestinal Tract
Animals

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Evaluation of Gram-negative bacterial infection by a stable and conjugative bioluminescence plasmid in a mouse model. / Huang, Yao Kuang; Chu, Chishih; Wu, Chih Hsiung; Chen, Chyi Liang; Chiu, Cheng Hsun.

In: Journal of Biomedical Science, Vol. 21, 2014, p. 78.

Research output: Contribution to journalArticle

Huang, Yao Kuang ; Chu, Chishih ; Wu, Chih Hsiung ; Chen, Chyi Liang ; Chiu, Cheng Hsun. / Evaluation of Gram-negative bacterial infection by a stable and conjugative bioluminescence plasmid in a mouse model. In: Journal of Biomedical Science. 2014 ; Vol. 21. pp. 78.
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AU - Huang, Yao Kuang

AU - Chu, Chishih

AU - Wu, Chih Hsiung

AU - Chen, Chyi Liang

AU - Chiu, Cheng Hsun

PY - 2014

Y1 - 2014

N2 - BACKGROUND: The green fluorescence protein (GFP)-associated fluorescence method and the luciferase-associated bioluminescence method are the two major methods for IVIS imaging system to investigate the bacterial infection in animal models. The aim of this study was to evaluate the infection route of Gram-negative bacteria carrying a stable and broad range of conjugative bioluminescence plasmid pSE-Lux1 in a mouse model.RESULTS: Both encapsulated and non-encapsulated Gram-negative bacteria were used as hosts to evaluate conjugation efficiency and plasmid stability of pSE-Lux1, a recombinant of pSE34 and luxABCDE operon. The plasmid conjugation efficiencies of pSE-Lux1 ranged from 10⁻³ to 10⁻⁷ in various Gram-negative bacteria. Plasmid pSE-Lux1 maintained in Escherichia coli, Klebsiella pneumoniae, and Salmonella enterica serovars Choleraesues (abbreviated S. Choleraesuis) and Typhimurium (S. Typhimurium), than in Acinetobacter baumannii and Serratia marcescens, was shown to be of better stability for at least four days. To investigate systemic bacterial infections, K. pneumoniae strain CG354 was intravenously injected, and then was clearly observed to be non-pathogenic to Balb/c mice for a long-term bioluminescence monitoring for 6 days. For examining dynamic distributions of gastrointestinal tract infection, the invasion protein SipB-deficient mutant OU5045△sipB and OU5046△sipB of S. serovar Typhimurium constructed in this study, compared to wild-type strain OU5045 and its virulence plasmid-less strain OU5046, were of less virulence to mice.CONCLUSIONS: This is the first study to evaluate the conjugative and stable bioluminescence vehicle system of pSE-Lux1 in a wide range of Gram-negative bacteria, a system that can provide a useful reporter approach to trace systemic and gastrointestinal bacterial infections in a mouse model.

AB - BACKGROUND: The green fluorescence protein (GFP)-associated fluorescence method and the luciferase-associated bioluminescence method are the two major methods for IVIS imaging system to investigate the bacterial infection in animal models. The aim of this study was to evaluate the infection route of Gram-negative bacteria carrying a stable and broad range of conjugative bioluminescence plasmid pSE-Lux1 in a mouse model.RESULTS: Both encapsulated and non-encapsulated Gram-negative bacteria were used as hosts to evaluate conjugation efficiency and plasmid stability of pSE-Lux1, a recombinant of pSE34 and luxABCDE operon. The plasmid conjugation efficiencies of pSE-Lux1 ranged from 10⁻³ to 10⁻⁷ in various Gram-negative bacteria. Plasmid pSE-Lux1 maintained in Escherichia coli, Klebsiella pneumoniae, and Salmonella enterica serovars Choleraesues (abbreviated S. Choleraesuis) and Typhimurium (S. Typhimurium), than in Acinetobacter baumannii and Serratia marcescens, was shown to be of better stability for at least four days. To investigate systemic bacterial infections, K. pneumoniae strain CG354 was intravenously injected, and then was clearly observed to be non-pathogenic to Balb/c mice for a long-term bioluminescence monitoring for 6 days. For examining dynamic distributions of gastrointestinal tract infection, the invasion protein SipB-deficient mutant OU5045△sipB and OU5046△sipB of S. serovar Typhimurium constructed in this study, compared to wild-type strain OU5045 and its virulence plasmid-less strain OU5046, were of less virulence to mice.CONCLUSIONS: This is the first study to evaluate the conjugative and stable bioluminescence vehicle system of pSE-Lux1 in a wide range of Gram-negative bacteria, a system that can provide a useful reporter approach to trace systemic and gastrointestinal bacterial infections in a mouse model.

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