转cry1Ac基因抗虫棉鄂杂棉1号的旁侧序列和品系特异性检测

摘要

To better understand the transformation events of transgenic cottons in China and to develope the rapid and accurate identification methods, the integrated structure and genomic flanking sequence of transgenic cotton (Gossypium hirsutum L.) Ezamian 1, the new event of transgenic insect-resistant cotton in china were identified using the long distance PCR (LD-PCR) and the genome walking methods.The event-specific qualitative PCR method for Ezamianl was established based on the flanking sequence through the optimization of primers, testing of the specificity and the sensitivity.The sensitivity of the method was 40 copies, corresponding to 0.1％ in 100 ng template.Twenty one samples from the market was tested with this method, three samples were identified having the same event structure of the Ezamianl.All of these indicated that the event-specific detection method established this study may be useful for the identification oftransgenic cotton in China.%为更好地了解我国转基因棉花的转化品系,建立快速准确的品系鉴别方法,本研究采用长链PCR、Genome Walking的方法解析转基因抗虫棉(Gossypium hirsutum L.)新转化品系鄂杂棉1号的整合结构和外源DNA整合的基因组旁侧序列.以旁侧序列为靶标,设计引物,对引物的筛选优化、特异性和灵敏度测试结果显示,建立的鄂杂棉1号品系定性PCR检测方法具有特异性,检测灵敏度为40个拷贝,相当于100 ng模板的0.1%.对市场的21个样品棉花样品检测,有3个样品检测出具有与"鄂杂棉1号"相同转化品系来源,表明本研究建立的品系特异性检测方法可应用于我国转基因棉花育种材料的鉴别和筛选.