Document Detail


DRESSA: biosensing of dioxin and dioxin-like chemicals using secreted alkaline phosphatase.
MedLine Citation:
PMID:  15519573     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In this article, we describe a highly sensitive biosensing system, DRESSA, for detection of dioxin and dioxin-like chemicals. Tandem copies of the dioxin-responsive element (DRE) fused to a minimal viral promoter were subcloned into an expression plasmid upstream of a secreted alkaline phosphatase (SEAP) gene. When murine hepatoma cell line Hepa-1c1c7 was stably transfected with this construct, established sensor clones secreted SEAP following stimulation with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). A clone HeDS49 was found to be extremely sensitive; it secreted SEAP in response to TCDD in dose- and time-dependent manners, and the minimal detection limit was 100 fM. To detect more than 6 pM of TCDD, the whole assay time (from cell seeding to measurement of SEAP activity) could be reduced to 4h. Secretion of SEAP was induced selectively by other activators of DRE (3-methylcholanthrene, benzo[a]pyrene, and beta-naphthoflavone) but not by activators of unrelated responsive elements. These data suggested that because of the rapidity, easiness, specificity, and high sensitivity of DRESSA, it is more suitable than currently available detection systems for dioxin and dioxin-like chemicals and would be of great advantage to high-throughput screening of these pollutants in environmental samples.
Authors:
Ayumi Kasai; Nobuhiko Hiramatsu; Yiman Meng; Jian Yao; Masayuki Takeda; Shuichiro Maeda; Masanori Kitamura
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical biochemistry     Volume:  335     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2004 Dec 
Date Detail:
Created Date:  2004-11-02     Completed Date:  2005-03-17     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  73-80     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Tamaho, Yamanashi 409-3898, Japan.
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MeSH Terms
Descriptor/Qualifier:
Alkaline Phosphatase / genetics,  secretion*
Animals
Benzo(a)pyrene / pharmacology
Carcinoma, Hepatocellular / metabolism,  pathology
Cytochrome P-450 CYP1A1 / genetics
Dioxins / pharmacology*
Enzyme Induction / drug effects
Gene Expression Regulation / drug effects*
Liver Neoplasms / metabolism,  pathology
Mammary Tumor Virus, Mouse / genetics
Methylcholanthrene / pharmacology
Mice
Plasmids
Promoter Regions, Genetic / genetics
Response Elements / physiology*
Sensitivity and Specificity
Tetrachlorodibenzodioxin / pharmacology
Transfection
Tumor Cells, Cultured
beta-Naphthoflavone / pharmacology
Chemical
Reg. No./Substance:
0/Dioxins; 1746-01-6/Tetrachlorodibenzodioxin; 50-32-8/Benzo(a)pyrene; 56-49-5/Methylcholanthrene; 6051-87-2/beta-Naphthoflavone; EC 1.14.14.1/Cytochrome P-450 CYP1A1; EC 3.1.3.1/Alkaline Phosphatase

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