Document Detail


Reporter gene assay for detection of shellfish toxins.
MedLine Citation:
PMID:  20163067     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
OBJECTIVE: To explore the potential reporter gene assay for the detection of sodium channel-specific toxins in shellfish as an alternative for screening harmful algal bloom (HAB) toxins, considering the fact that the existing methods including HPLC and bioassay are inappropriate for identifying HAB toxins which poses a serious problem on human health and shellfish industry. METHODS: A reporter plasmid pEGFP-c-fos containing c-fos promoter and EGFP was constructed and transfected into T24 cells using LipofectAMINE 2000. Positive transfectants were screened by G418 to produce a pEGFP-c-fos-T24 cell line. After addition of increasing neurotoxic shellfish poison (NSP) or GTX2,3, primary components of paralytic shellfish poison (PSP), changes in expression of EGFP in the cell line were observed under a laser scanning confocal microscope and quantified with Image-pro Plus software. RESULTS: Dose-dependent changes in the intensity of green fluorescence were observed for NSP in a range from 0 to 10 ng/mL and for GTX2,3 from 0 to 16 ng/mL. CONCLUSION: pEGFP-c-fos-T24 can be applied in detecting HAB toxins, and cell-based assay can be used as an alternative for screening sodium channel-specific HAB toxins.
Authors:
Wei-Dong Yang; Min-Yi Wu; Jie-Sheng Liu; Xi-Chun Peng; Hong-Ye Li
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biomedical and environmental sciences : BES     Volume:  22     ISSN:  0895-3988     ISO Abbreviation:  Biomed. Environ. Sci.     Publication Date:  2009 Oct 
Date Detail:
Created Date:  2010-02-18     Completed Date:  2010-03-16     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8909524     Medline TA:  Biomed Environ Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  419-22     Citation Subset:  IM    
Affiliation:
School of Life Science and Technology, Jinan University, Guangzhou 510632, Guangdong, China. tywd@jnu.edu.cn
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Assay
Cell Line, Tumor
Genes, Reporter / physiology*
Green Fluorescent Proteins
Harmful Algal Bloom / physiology
Humans
Plasmids
Proto-Oncogene Proteins c-fos / genetics,  metabolism
Shellfish / analysis*
Sodium Channels
Toxins, Biological / chemistry*,  toxicity*
Chemical
Reg. No./Substance:
0/Proto-Oncogene Proteins c-fos; 0/Sodium Channels; 0/Toxins, Biological; 147336-22-9/Green Fluorescent Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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