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

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.

journal_name

FEMS Microbiol Ecol

authors

Ramaiah N,Chun J,Ravel J,Straube WL,Hill RT,Colwell RR

doi

10.1111/j.1574-6941.2000.tb00723.x

subject

Has Abstract

pub_date

2000-07-01 00:00:00

pages

27-34

issue

1

eissn

0168-6496

issn

1574-6941

pii

S0168-6496(00)00040-4

journal_volume

33

pub_type

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