Screening Detection of Bt Genes in Genetically Modified Crops Using a Multiplex PCR Assay
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Abstract:
Since Bt genes were widely used in insect-resistant transgenic crops, they became the most important target genes in screening genetically modified food. Based on nucleotide sequences analysis, eight common Bt genes (cry1Ab, cry1Ac, cry1Ab/Ac, cry1A.105, cry1Ac-M, cry2Ab, cry3A and cry3Bb) in transgenic crops were divided into cry1A, cry2A and cry3A groups. To detect three groups of Bt genes, degenerate primers for cry1A group and specific primers for cry2A and cry3A groups were designed according to the consensus sequences of each group, and three singlet PCR methods for each group respectively were established after testing specificity and sensitivity in this study. In addition, using the three pairs of PCR primers, a triplex PCR method also established to screen the three groups of Bt genes simultaneously. The results revealed that these methods could accurately detect the targeted Bt genes from the various transgenic samples, and the detection sensitivity was up to 0.1%. As the method established in this study showed highly specific and sensitive, it would be a good prospect in the screening of genetically modified organism.
