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花匠小妙招
2024-08-10 11:49

实时荧光RT-PCR一步法检测南方菜豆花叶病毒点此下载全文引用本文：白静,陈红运,傅俊范,李桂芬,朱水芳.实时荧光RT-PCR一步法检测南方菜豆花叶病毒.植物保护学报,2007,34(1):78-82 DOI：摘要点击次数:1318 全文下载次数:340 作者单位白静沈阳农业大学植物保护学院,沈阳 110161
中国检验检疫科学研究院动植物检疫研究所,北京 100029 陈红运中国检验检疫科学研究院动植物检疫研究所,北京 100029 傅俊范沈阳农业大学植物保护学院,沈阳 110161 李桂芬中国检验检疫科学研究院动植物检疫研究所,北京 100029 朱水芳中国检验检疫科学研究院动植物检疫研究所,北京 100029 中文摘要:南方菜豆花叶病毒(Southern bean mosaic virus,SBMV)是我国二类检疫性有害生物,以南方菜豆花叶病毒日本分离物(SBMV-J)总RNA为模板,采用RT-PCR方法扩增病毒外壳蛋白基因及其上游基因的cDNA片断并将其克隆到pMD18-T载体上。序列分析结果表明:SBMV-J cp基因由801个核苷酸组成,编码266个氨基酸,SBMV-J与其它分离物及株系cp基因的核苷酸序列同源性为83% ~97％,氨基酸序列同源性为86% ~97％。由于SBMV各分离物及株系cp基因的同源性较低,难于设计出较长的普通PCR引物。通过较短引物设计和TaqMan-MGB探针技术,建立了SBMV的实时荧光RT-PCR一步检测方法。该方法的检测低限是0.16pg,最佳检测总RNA的量是0.16ng。中文关键词:南方菜豆花叶病毒外壳蛋白基因序列分析实时荧光RT-PCR Detection of Southern bean mosaic virus by one step assay of real-time fluorescent RT-PCR Author NameAffiliationBAI Jing Department of Plant Protection, Shenyang Agricultural University, Shenyang 110161, Liaoning Province, China
Institute of Animal and Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China CHEN Hong-yun Institute of Animal and Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China FU Jun-fan Department of Plant Protection, Shenyang Agricultural University, Shenyang 110161, Liaoning Province, China LI Gui-fen Institute of Animal and Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China ZHU Shui-fang Institute of Animal and Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China Abstract:Southern bean mosaic virus (SBMV) is in the A2 list concerned to China. The coding region of cp gene from Japan isolate of SBMV was sequenced. It comprised 801 nucleotides and encoded a putative protein of 266 amino acids. The nucleotide and amino acid sequence of cp gene of this isolate shared 83% to 97% and 86% to 97% similarities to those of other SBMV isolates or strains, respectively. A pair of primers and a TaqMan-MGB probe based on the conserved nucleotide sequence of coat protein (CP) gene of SBMV isolates were designed and synthesized. A novel one step assay of real-time fluorescent RT-PCR was established to detect SBMV. The limit content of the detection was 0.16pg and the optimal content was 0.16ng.keywords:Southern bean mosaic virus (SBMV) cp gene sequence analysis TaqMan-MGB probe 查看全文查看/发表评论下载PDF阅读器

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