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等位基因特异性扩增(Allele-specific PCR简称:ASPCR)技术是一项快速、简便、低成本的检测等位基因单核苷酸多态性的新方法。作者针对雨久花感抗磺酰脲类生态型ALS基因编码区第592位碱基密码(ALS氨基酸第197位)特征,采用在引物3′末端设有胸腺嘧啶核苷酸(感性)或鸟嘌呤(抗性)核苷酸的2种引物,对延边稻田发生的雨久花感抗磺酰脲类生态型ALS基因编码区第592位进行了ASPCR。检测结果显示,当以引物3′末端设有胸腺嘧啶引物时,感性类型出现ASPCR扩增带,而抗性类型无带出现,反之抗性类型有带而感性类型无带,检测到延边稻田发生的抗磺酰脲类雨久花生态型的ALS 197位点的脯氨酸被组氨酸替代,证明了该方法的有效性和可靠性。
Allele-specific PCR (ASPCR) is a rapid, simple and low-cost method to detect single nucleotide polymorphism of alleles. The aim of this study is to characterize the 591 base pair codon (amino acid 197 of ALS) in the coding sequence of the sulfonylurea-resistant ALS gene in Yucaoxuyin, using thymine nucleotide (inducing) or 3 Purine (resistant) nucleotides. The results showed that ASPCR was the 592th position in the coding region of the ALS gene of sulfonylurea, which occurred in Yanbian paddy field. The results showed that when the thymidine primer was located at the 3 ’end of the primer, the ASPCR amplification band appeared in the inductive type, but the band of the resistant type appeared without the band, whereas the band of the resistant type and the band of the inductive type did not appear, and the detection of the paddy field in Yanbian occurred Of the proline of the ALS 197 site of the sulfonylurea-type long-flowering ecotype was replaced by histidine, demonstrating the effectiveness and reliability of the method.