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Detection of luciferase gene sequences in nonluminescent bacteria from the Chesapeake Bay

Cited 16 time in Web of Science Cited 16 time in Scopus
Authors
Ramaiah, Nahappa; Chun, Jongsik; Ravel, Jacques; Straube, William L; Hill, Russell T; Colwell, Rita R
Issue Date
2000-07
Citation
FEMS Microbiology Ecology, Vol.33 No.1, pp.27-34
Keywords
bioluminescent bacterialuxA geneChesapeake BayDNA probe
Abstract
A 745-bp luxA fragment was amplified from Vibrio harveyi (UM 1503), radiolabeled, and used as a probe to detect and quantify luxA genotypes in culturable bacterial populations from the Chesapeake Bay. DNA samples from 53 reference strains were also examined for this gene. The luxA-positive bacteria comprised from 0-6% of the culturable heterotrophic bacterial community in samples from the Bay. Only those reference strains known to be luminescent contained the luxA gene, as indicated by PCR. Results in all cases were confirmed by PCR of DNA extracts and Southern hybridization analyses, using an internal probe for confirmation of luxA amplification products. Sequence analysis of luxA genes from three nonluminescent bacteria isolated from the Chesapeake Bay indicated little or no differences when compared with luxA sequences from known marine luminescent bacterial species. These three Chesapeake Bay strains and other luxA- positive strains were tested with a luminometer and confirmed to be nonluminescent. All of over 7800 bacterial colonies enumerated during this study from Chesapeake Bay samples were non-visibly luminescent. Our results indicate that luxA-positive bacteria isolated from the Chesapeake Bay are not generally luminescent on phenotypic examination, implying that gene probe techniques are required for examining luxA gene distribution in microbial populations present in environmental samples. (C) 2000 Published by Elsevier Science B.V. on behalf of the Federation of European Microbiological Societies.
ISSN
0168-6496
URI
http://hdl.handle.net/10371/166168
DOI
https://doi.org/10.1016/S0168-6496(00)00040-4
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College of Natural Sciences (자연과학대학)Dept. of Biological Sciences (생명과학부)Journal Papers (저널논문_생명과학부)
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