Development of species-specific hybridization probes for marine luminous bacteria by using in vitro DNA amplification

Journal Article · Wed May 01 04:00:00 EDT 1991 · Applied and Environmental Microbiology; (United States)

OSTI ID:5426982

Wimpee, C F; Nadeau, T L; Nealson, K H ^[1]

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1. Univ. of Wisconsin, Milwaukee (USA)

By using two highly conserved regions of the luxA gene as primers, polymerase chain reaction amplification methods were used to prepare species-specific probes against the luciferase gene from four major groups of marine luminous bacteria. Laboratory studies with test strains indicated that three of the four probes cross-reacted with themselves and with one or more of the other species at low stringencies but were specific for members of their own species at high stringencies. The fourth probe, generated from Vibrio harveyi DNA, a cross-reacted with DNAs from two closely related species, V. orientalis and V. vulnificus. When nonluminous cultures were tested with the species-specific probes, no false-positive results were observed, even at low stringencies. Two field isolates were correctly identified as Photobacterium phosphoreum by using the species-specific hybridization probes at high stringency. A mixed probe (four different hybridization probes) used at low stringency gave positive results with all of the luminous bacteria tested, including the terrestrial species Xenorhabdus luminescens, and the taxonomically distinct marine bacterial species Shewanella hanedai; minimal cross-hybridization with these species was seen at higher stringencies.

• OSTI ID: 5426982
• Journal Information: Applied and Environmental Microbiology; (United States), Journal Name: Applied and Environmental Microbiology; (United States) Vol. 57:5; ISSN 0099-2240; ISSN AEMID
• Country of Publication: United States
• Language: English

Related Subjects

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54 ENVIRONMENTAL SCIENCES
540310 -- Environment
Aquatic-- Basic Studies-- (1990-)
BACTERIA
BIOLOGY
BIOLUMINESCENCE
BIOTECHNOLOGY
DNA HYBRIDIZATION
DNA POLYMERASES
ENZYMES
GENE AMPLIFICATION
GENES
GENETIC ENGINEERING
HYBRIDIZATION
IN VITRO
LUCIFERASE
LUMINESCENCE
MICROORGANISMS
NUCLEOTIDYLTRANSFERASES
OXIDOREDUCTASES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
SEAS
SURFACE WATERS
TAXONOMY
TRANSFERASES