L5 luciferase reporter mycobacteriophages: a sensitive tool for the detection and assay of live mycobacteria.
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.
Sarkis
G J
GJ
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260, USA.
Jacobs
W R
WR
Hatfull
G F
GF
D015373:Bacterial Typing Techniques; D001483:Base Sequence; D004352:Drug Resistance, Microbial; D004977:Ethambutol / Q000494:pharmacology; D017344:Genes, Insect / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral; D007538:Isoniazid / Q000494:pharmacology; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008242:Lysogeny; D008969:Molecular Sequence Data; D009160:Mycobacteriophages / Q000235:genetics / Q000254:growth & development; D009161:Mycobacterium / Q000145:classification / Q000187:drug effects / Q000302:isolation & purification / Q000821:virology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012293:Rifampin / Q000494:pharmacology; D012680:Sensitivity and Specificity; D013307:Streptomycin / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D004977:Ethambutol; D008156:Luciferases; D007538:Isoniazid; D012293:Rifampin; D013307:Streptomycin