A REPORTER GENE ASSAY FOR THE DETECTION OF SHELLFISH TOXINS

来源 :第十四届世界食品科技大会 | 被引量 : 0次 | 上传用户:kaixun520
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  A reporter plasmid pEGFP-c-fos containing c-los promoter and enhanced green fluorescent protein (EGFP) was constructed and transfected into T24 cell using LipofectAMINE 2000.The positive transfectants were screened by G418 to produce a pEGFP-c-fos-T24 cell line.Changes in expression of EGFP in the cell line after addition of increasing neurotoxic shellfish poison (NSP) or GTX2,3,primary components of paralytic shellfish poison (PSP),were detected using a laser scanning confocal microscope and quantified with Image-pro Plus software.Dose-dependent changes of the intensity of green fluorescence were observed for NSP in a range of 0 ~10 ng/ml and for GTX2,3 from 0 to 16 ng/ml.These results supported the applicability of pEGFP-c-fos-T24 in the detection of harmful algal blooms (HABs) toxins in shellfish or algae.Therefore,this cell-based assay provides an alternative for the screening of sodium channel-specific HABs toxins.
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