Abstract:
:A simple, rapid, sensitive microtiter plate method detecting N-acyl homoserine lactone (HSL)-related compounds was established using an Agrobacterium tumefaciens strain harboring a traG::lacZ/traR reporter gene responsive to HSLs. This strain did not produce its own HSL, but the traG::lacZ reporter gene was induced only when its transcription activator TraR detected a cognate exogenous HSL. Therefore, the assay was expected to be highly specific for HSL-related compounds. Induction of the reporter gene, leading to production of beta-galactosidase enzyme, was measured by using two different beta-galactosidase substrates, X-gal and Galacton-Star, for colorimetric and chemiluminometric detection, respectively. The screen was validated in both the 96-well and 384-well plate formats, and extracts derived from 696 different microbial isolates, mostly unidentified actinomycetes isolated from diverse locations, were tested. Crude extracts of 81 (11.64%) cultures tested positive for HSL-related compounds, and an additional 34 (4.8%) crude extracts showed a moderate to weak signal for HSLs. Data from the fractionated samples, however, suggested a much higher prevalence of HSL signals in these extracts. Of 144 crude extracts fractionated into 10 individual samples at a 10x concentration, 72 (50%) cultures tested positive for HSLs. Six cultures were active only in the crude extract, 18 were active both in crude and one or more of their fractions, and an additional 48 were active in just one or more of their fractions. This finding may be the first to suggest such a high prevalence of HSL-signals found in nature, and a large number of actinomycetes in our collection appeared to produce HSL-related compounds.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Singh MP,Greenstein Mdoi
10.1016/j.mimet.2005.06.003subject
Has Abstractpub_date
2006-04-01 00:00:00pages
32-7issue
1eissn
0167-7012issn
1872-8359pii
S0167-7012(05)00163-6journal_volume
65pub_type
杂志文章abstract::In this study, we aimed to identify the molecular carbapenemase types of the Enterobacteriaceae isolates and to evaluate the performance of manually prepared and commercially available combination disc methods and the modified Hodge test. One hundred and forty carbapenemase producing isolates and 45 isolates as contro...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
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journal_title:Journal of microbiological methods
pub_type: 杂志文章,评审
doi:10.1016/j.mimet.2004.12.003
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journal_title:Journal of microbiological methods
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journal_title:Journal of microbiological methods
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doi:10.1016/j.mimet.2007.05.016
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journal_title:Journal of microbiological methods
pub_type: 杂志文章
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journal_title:Journal of microbiological methods
pub_type: 杂志文章,多中心研究
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journal_title:Journal of microbiological methods
pub_type: 杂志文章
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pub_type: 杂志文章
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journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.01.003
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更新日期:2015-09-01 00:00:00
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journal_title:Journal of microbiological methods
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journal_title:Journal of microbiological methods
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journal_title:Journal of microbiological methods
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journal_title:Journal of microbiological methods
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