| pubmed-article:18424536 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:18424536 | lifeskim:mentions | umls-concept:C0220908 | lld:lifeskim |
| pubmed-article:18424536 | lifeskim:mentions | umls-concept:C1710082 | lld:lifeskim |
| pubmed-article:18424536 | lifeskim:mentions | umls-concept:C1533691 | lld:lifeskim |
| pubmed-article:18424536 | lifeskim:mentions | umls-concept:C1510438 | lld:lifeskim |
| pubmed-article:18424536 | lifeskim:mentions | umls-concept:C0439831 | lld:lifeskim |
| pubmed-article:18424536 | lifeskim:mentions | umls-concept:C0257136 | lld:lifeskim |
| pubmed-article:18424536 | pubmed:issue | 12 | lld:pubmed |
| pubmed-article:18424536 | pubmed:dateCreated | 2008-6-16 | lld:pubmed |
| pubmed-article:18424536 | pubmed:abstractText | A simple, sensitive, and rapid cell-free assay system was developed for detection of N-acyl homoserine lactone (AHL) autoinducers involved in bacterial quorum sensing (QS). The present approach improves upon previous whole-cell biosensor-based approaches in its utilization of a cell-free assay approach to conduct bioassays. The cell-free assay was derived from the AHL biosensor bacterium Agrobacterium tumefaciens NTL4(pCF218)(pCF372), allowing the expression of beta-galactosidase upon addition of exogenous AHLs. We have shown that beta-galactosidase expression is possible in cell-free solution [lysate from Agrobacterium tumefaciens NTL4(pCF218)(pCF372) culture]. Assay detection limits with the use of chromogenic substrate X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) ranged from approximately 100 nM to 300 nM depending on the specific AHL. Replacement (of X-Gal) with the luminescent substrate Beta-Glo increased sensitivity to AHLs by 10-fold. A major advantage of the cell-free assay system is elimination of time-consuming steps for biosensor cell culture conditioning, which are required prior to whole-cell bioassays. This significantly reduced assay times from greater than 24 h to less than 3 h, while maintaining high sensitivity. Assay lysate may be prepared in bulk and stored (-80 degrees C) over 6 months for future use. Finally, the present protocol may be adapted for use with other biosensor strains and be used in high-throughput AHL screening of bacteria or metagenomic libraries. | lld:pubmed |
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| pubmed-article:18424536 | pubmed:language | eng | lld:pubmed |
| pubmed-article:18424536 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:18424536 | pubmed:citationSubset | IM | lld:pubmed |
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| pubmed-article:18424536 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:18424536 | pubmed:month | Jun | lld:pubmed |
| pubmed-article:18424536 | pubmed:issn | 1098-5336 | lld:pubmed |
| pubmed-article:18424536 | pubmed:author | pubmed-author:KawaguchiTomo... | lld:pubmed |
| pubmed-article:18424536 | pubmed:author | pubmed-author:DechoAlan WAW | lld:pubmed |
| pubmed-article:18424536 | pubmed:author | pubmed-author:NormanR... | lld:pubmed |
| pubmed-article:18424536 | pubmed:author | pubmed-author:ChenYung... | lld:pubmed |
| pubmed-article:18424536 | pubmed:issnType | Electronic | lld:pubmed |
| pubmed-article:18424536 | pubmed:volume | 74 | lld:pubmed |
| pubmed-article:18424536 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:18424536 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:18424536 | pubmed:pagination | 3667-71 | lld:pubmed |
| pubmed-article:18424536 | pubmed:dateRevised | 2011-11-17 | lld:pubmed |
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| pubmed-article:18424536 | pubmed:meshHeading | pubmed-meshheading:18424536... | lld:pubmed |
| pubmed-article:18424536 | pubmed:year | 2008 | lld:pubmed |
| pubmed-article:18424536 | pubmed:articleTitle | Rapid screening of quorum-sensing signal N-acyl homoserine lactones by an in vitro cell-free assay. | lld:pubmed |
| pubmed-article:18424536 | pubmed:affiliation | Department of Environmental Health Sciences, Arnold School of Public Health, University of South Carolina, Columbia, SC 29208, USA. kawaguch@gwm.sc.edu | lld:pubmed |
| pubmed-article:18424536 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:18424536 | pubmed:publicationType | Comparative Study | lld:pubmed |
| pubmed-article:18424536 | pubmed:publicationType | Research Support, U.S. Gov't, Non-P.H.S. | lld:pubmed |
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