L5 luciferase reporter mycobacteriophages: a sensitive tool for the detection and assay of live mycobacteria

Gary J. Sarkis; William R. Jacobs; Graham F. Hatfulll

doi:10.1111/j.1365-2958.1995.tb02281.x

L5 luciferase reporter mycobacteriophages: a sensitive tool for the detection and assay of live mycobacteria

Gary J. Sarkis, William R. Jacobs, Graham F. Hatfulll

Microbiology & Immunology

Research output: Contribution to journal › Article › peer-review

122 Scopus citations

Abstract

Recombinant bacteriophages provide efficient delivery systems for introducing reporter genes into specific bacterial hosts. We have constructed mycobacteriophage L5 recombinants carrying the firefly luciferase gene inserted into the tRNA region of the phage genome. Infection of Mycobacterium smegmatis by these phages results in expression of the luciferase gene and light emission. Fortuitously, the luciferase gene is expressed continuously in lysogens surviving infection. Synthesis of luciferase from a mycobacterial promoter created by cloning enables the detection of extremely small numbers of M. smegmatis cells. These reporter phages can be used to discriminate between drug‐sensitive and drug‐resistant strains of M. smegmatis, and may provide tools for the rapid identification and classification of antimycobacterial agents.

Original language	English (US)
Pages (from-to)	1055-1067
Number of pages	13
Journal	Molecular Microbiology
Volume	15
Issue number	6
DOIs	https://doi.org/10.1111/j.1365-2958.1995.tb02281.x
State	Published - Mar 1995

ASJC Scopus subject areas

Microbiology
Molecular Biology

Access to Document

10.1111/j.1365-2958.1995.tb02281.x

Cite this

@article{0801bd61744948829a0d6365287f58dd,

title = "L5 luciferase reporter mycobacteriophages: a sensitive tool for the detection and assay of live mycobacteria",

abstract = "Recombinant bacteriophages provide efficient delivery systems for introducing reporter genes into specific bacterial hosts. We have constructed mycobacteriophage L5 recombinants carrying the firefly luciferase gene inserted into the tRNA region of the phage genome. Infection of Mycobacterium smegmatis by these phages results in expression of the luciferase gene and light emission. Fortuitously, the luciferase gene is expressed continuously in lysogens surviving infection. Synthesis of luciferase from a mycobacterial promoter created by cloning enables the detection of extremely small numbers of M. smegmatis cells. These reporter phages can be used to discriminate between drug‐sensitive and drug‐resistant strains of M. smegmatis, and may provide tools for the rapid identification and classification of antimycobacterial agents.",

author = "Sarkis, {Gary J.} and Jacobs, {William R.} and Hatfulll, {Graham F.}",

year = "1995",

month = mar,

doi = "10.1111/j.1365-2958.1995.tb02281.x",

language = "English (US)",

volume = "15",

pages = "1055--1067",

journal = "Molecular Microbiology",

issn = "0950-382X",

publisher = "Wiley-Blackwell",

number = "6",

}

TY - JOUR

T1 - L5 luciferase reporter mycobacteriophages

T2 - a sensitive tool for the detection and assay of live mycobacteria

AU - Sarkis, Gary J.

AU - Jacobs, William R.

AU - Hatfulll, Graham F.

PY - 1995/3

Y1 - 1995/3

N2 - Recombinant bacteriophages provide efficient delivery systems for introducing reporter genes into specific bacterial hosts. We have constructed mycobacteriophage L5 recombinants carrying the firefly luciferase gene inserted into the tRNA region of the phage genome. Infection of Mycobacterium smegmatis by these phages results in expression of the luciferase gene and light emission. Fortuitously, the luciferase gene is expressed continuously in lysogens surviving infection. Synthesis of luciferase from a mycobacterial promoter created by cloning enables the detection of extremely small numbers of M. smegmatis cells. These reporter phages can be used to discriminate between drug‐sensitive and drug‐resistant strains of M. smegmatis, and may provide tools for the rapid identification and classification of antimycobacterial agents.

AB - Recombinant bacteriophages provide efficient delivery systems for introducing reporter genes into specific bacterial hosts. We have constructed mycobacteriophage L5 recombinants carrying the firefly luciferase gene inserted into the tRNA region of the phage genome. Infection of Mycobacterium smegmatis by these phages results in expression of the luciferase gene and light emission. Fortuitously, the luciferase gene is expressed continuously in lysogens surviving infection. Synthesis of luciferase from a mycobacterial promoter created by cloning enables the detection of extremely small numbers of M. smegmatis cells. These reporter phages can be used to discriminate between drug‐sensitive and drug‐resistant strains of M. smegmatis, and may provide tools for the rapid identification and classification of antimycobacterial agents.

UR - http://www.scopus.com/inward/record.url?scp=0028940695&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028940695&partnerID=8YFLogxK

U2 - 10.1111/j.1365-2958.1995.tb02281.x

DO - 10.1111/j.1365-2958.1995.tb02281.x

M3 - Article

C2 - 7623662

AN - SCOPUS:0028940695

SN - 0950-382X

VL - 15

SP - 1055

EP - 1067

JO - Molecular Microbiology

JF - Molecular Microbiology

IS - 6

ER -

L5 luciferase reporter mycobacteriophages: a sensitive tool for the detection and assay of live mycobacteria

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this