用特异引物扩增核糖体DNA鉴定咖啡扇叶螨和松叶螨

T. Uehara, T. Mizukubo, A. Kushida, Y. Momota
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引用次数: 51

摘要

用PCR方法从基因组DNA中扩增出咖啡和松孢霉rDNA的18-26S区。序列分析表明,58s基因序列具有保守性,而其内部转录间隔物(ITS)序列具有分化性。利用ITS序列差异合成特异性引物,区分咖啡假蝇和松蝇。用这些引物特异性扩增每个物种的特征DNA片段。此外,从单个雌虫、雄虫或幼虫中提取的DNA也获得了相同的特异性扩增产物。
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Identification of Pratylenchus Coffeae and P. Loosi Using Specific Primers for PCR Amplification of Ribosomal DNA
The 18-26S region of rDNA of P. coffeae and P. loosi were amplified by PCR from genomic DNA. Sequence analyses indicated that the 5.8S gene sequences were conserved whereas the internal transcribed spacer (ITS) sequences were divergent. Sequence differences in the ITS were used to synthesize specific primer sets to differentiate P. coffeae and P. loosi. PCR assays with these primers specifically amplified a characteristic DNA fragment from each species. Moreover, the same specific amplification products were obtained using DNA extracted from single females males or juveniles.
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