生物技术通报 ›› 2020, Vol. 36 ›› Issue (5): 39-47.doi: 10.13560/j.cnki.biotech.bull.1985.2019-1090

• 生物分析方法与标准物质专题(专题主编:李亮 副研究员) • 上一篇    下一篇

玉米转基因成分筛查策略

温洪涛1, 李夏莹2, 杨洋1, 陈子言2, 丁一佳1, 张秀杰2, 张瑞英1   

  1. 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
  • 收稿日期:2019-11-11 出版日期:2020-05-26 发布日期:2020-06-03
  • 作者简介:温洪涛,男,博士,副研究员,研究方向:转基因检测与安全评价;E-mail:wen0891@163.com
  • 基金资助:
    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)

Strategy of Screening Genetically Modified Maize

WEN Hong-tao1, LI Xia-ying2, YANG Yang1, CHEN Zi-yan2, DING Yi-jia1, ZHANG Xiu-jie2, ZHANG Rui-ying1   

  1. 1. Quality & Safety Institute of Agricultural Products,Heilongjiang Academy of Agricultural Sciences,Harbin 150086;
    2. Development Center of Science and Technology,Ministry of Agriculture and Rural Affairs,Beijing 100176
  • Received:2019-11-11 Published:2020-05-26 Online:2020-06-03

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、NPT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。

关键词: 转基因玉米, 筛查, 转化体, 鉴定

Abstract: 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.

Key words: genetically modified maize, screening, GMO event, detection