耐除草剂玉米MON87427定性PCR检测方法的建立

Establishment of Qualitative PCR Detection Method forHerbicide-tolerant Maize MON87427

MON87427转化体是美国孟山都公司研发的转基因耐除草剂玉米,已经被包括我国在内的18个国家授权应用。对该转化体的快速检测是实现对其日常监管的基本前提。为了建立MON87427特异性的检测方法,研究针对其特征DNA序列筛选了最佳引物组合,并对其PCR反应过程进行优化,以初步建立转基因玉米MON87427转化体定性PCR检测方法。结果表明,经特异性测试,引物组合FA/RC能特异性检出MON87427玉米,对其他主要作物或转化体无效;该方法的检测灵敏度可达0.05%。研究建立的新方法可以满足我国现行转基因作物监管检测技术要求,适用于对MON87427转化体特异的定性PCR检测。

The MON87427 event is a genetically modified(GM)herbicide-resistant corn developed by Monsanto Monsanto Company in the United States and is authorized for use in 18 countries,including China.Qualitative testing of the event is a fundamental prerequisite for achieving its daily supervision.To establish a MON87427-specific detection method,this study screened the optimal combination of primers targeting its characteristic DNA sequence and optimized its PCR reaction process,to establish a qualitative PCR method for MON87427 event.The result showed that primer combination FA/RC was found to be specific for MON87427 maize and not for other major crops or GM transformants.And the limit of detection of this method could be up to 0.05%.These results show that the novel detection method can meet the technical requirements of the current regulatory testing of transgenic crops in China and is suitable for the specific PCR detection of MON87427 events qualitatively.