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KMID : 1007520080170020357
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Food Science and Biotechnology
2008 Volume.17 No. 2 p.357 ~ p.361
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A Duplex PCR Assay for Differentiating Native Common Buckwheat and Tartarian Buckwheat, and Its Application for the Rapid Detection of Buckwheat Ingredients in Food
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Jeon Young-Jun
Hong Kwang-Won
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Abstract
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One of the major allergenic proteins in common buckwheat (Fagopyrum elculentum) was found to be a BW10KD. In this work, allergenic BW10KD genomic DNAs from the native common buckwheat ``Pyeongchang`` and Tartarian buckwheat ``Clfa47`` were cloned by polymerase chain reaction (PCR), and their nucleotide sequences were determined. In addition, a novel PCR assay targeting the allergenic BW10KD gene was developed to detect and differentiate both buckwheat species in food. The nucleotide sequences of the BW10KD genomic DNA from ``Pyeongchang`` and ``Clfa47`` were 94% identical. Base differences in the nucleotide sequences of the BW10KD genes are probably useful as a molecular marker for species-specific identification. The ``Pyeongchang``-specific primer set 154PF/400PR and the ``Clfa47``-specific primer set 154DF/253DR generated 247 and 100 bp fragments in singleplex PCR, respectively. A duplex PCR assay with 2 species-specific primer sets simultaneously differentiated the ``Pyeongchang`` and ``Clfa47`` in a single reaction. The PCR assay also successfully allowed for the rapid detection of buckwheat ingredients in foods.
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KEYWORD
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common buckwheat, Tartarian buckwheat, food allergen, BW10KD, polymerase chain reaction, PCR
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