玉米转基因成分筛查策略  

Strategy of Screening Genetically Modified Maize

作　　者：温洪涛 李夏莹 杨洋 陈子言 丁一佳 张秀杰 张瑞英 WEN Hong-tao;LI Xia-ying;YANG Yang;CHEN Zi-yan;DING Yi-jia;ZHANG Xiu-jie;ZHANG Rui-ying(Quality&Safety Institute of Agricultural Products,Heilongjiang Academy of Agricultural Sciences,Harbin 150086;Development Center of Science and Technology,Ministry of Agriculture and Rural Affairs,Beijing 100176)

机构地区：[1]黑龙江省农业科学院农产品质量安全研究所,哈尔滨150086 [2]农业农村部科技发展中心,北京100176

出　　处：《生物技术通报》2020年第5期39-47,共9页Biotechnology Bulletin

基　　金：转基因新品种培育重大专项(2018ZX08011-04B);主要非授权转基因作物及其产品检测技术研究,黑龙江省农业科学院博士启动项目(201507-30)。

摘　　要：本研究通过查阅相关数据库中转基因玉米品系中的常用遗传转化载体中含有的调控元件和标记基因,并结合目前国内外现有转基因检测标准方法,建立了基于P-CaMV 35S、T-NOS、Bar基因、Pat基因、Cp4-epsps基因、NPTⅡ基因、cry1A基因、P-Rice actin、phy结构特异性片段等9种元件/基因实时荧光PCR方法的玉米转基因筛查策略,并对32种转基因玉米进行筛查实验测试,结果表明本筛查策略可覆盖30种独立性状转化体。同时还构建了包含玉米内标基因及9种目标元件/基因序列的质粒分子,经过验证该质粒分子具有良好的可替代性,可作为阳性对照与本检测方法配套使用。By checking regulatory elements and marker genes in common genetic transformation vectors of GM maize lines in relevant databases and combining it with the current domestic and foreign standard methods for the detection of GM,the strategy of screening transgenic maize by real-time fluorescent PCR method was established based on the P-CaMV35S,T-NOS,Bar gene,Pat gene,Cp4-epsps gene,NPT II gene,cry1A gene,P-Rice actin,and phy structure specific fragments,and 32 varieties of transgenic maize were tested in the screening.The results showed that the screening strategy covered 30 independent trait GMO events.Meanwhile,a plasmid molecule containing maize internal standard gene and 9 target elements/gene sequences was constructed.It was proved that the plasmid molecule had favorable substitution and could be used as a positive control in conjunction with this detection method.

关 键 词：转基因玉米 筛查 转化体 鉴定 

分 类 号：S513[农业科学—作物学]