转基因抗草甘膦油菜的检测方法.pdfVIP

生 物 技 术 通 讯 242 LETTERSINBIOTECHNOLOGY Vol.23No.2 Mar., 2012 doi: 10.3969/j.issn.1009-0002.2012.02.022 研究报告 转基因抗草甘膦油菜的检测方法 1 1 2 2 1 1 1 1 1 李闻娟 ，张建平 ，王少宏 ，孙佳 ，党占海 ，赵利 ，赵玮 ，王利民 ，党照 1. 甘肃省农业科学院 作物研究所，甘肃 兰州 730070；2. 甘肃农业大学，甘肃 兰州 730070 ［摘要］ 目的：建立检测田间转基因抗草甘膦油菜的方法。方法：用CTAB法提取DNA，经PCR分别扩增内参基因 BnACCg8 ，及抗草甘膦外源基因CP4-EPSPS 、FMV 35S启动子和E9 3终止子等4种基因的片段，以琼脂糖凝胶电泳 对扩增产物进行分析比对。结果：抗草甘膦油菜中分别扩增出与外源基因FMV 35S启动子、E9 3终止子和 CP4-EPSPS 大小一致的片段。结论：该方法能有效地用于转基因抗草甘膦油菜的检测。 ［关键词］ 抗草甘膦；转基因；油菜；PCR检测 ［中图分类号］ Q943.1 ［文献标识码］ A ［文章编号］ 1009-0002（2012）02-0242-03 A Method for Detection of Glyphosate Resistant Transgenic Rape 1 1 2 2 1 1 LI Wen-Juan , ZHANG Jian-Ping *, WANG Shao-Hong , SUN Jia , DANG Zhan-Hai , ZHAO Li , 1 1 1 ZHAO Wei , WANG Li-Min , DANG Zhao 1. Crops Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070; 2. Gansu Agricultural Uni⁃ versity, Lanzhou 730070; China *Corresponding author, E-mail: zhangjianpinglanzhou@ ［Abstract ］ Objective: To develop a method for detection of transgenic glyphosate resistant rape. Methods: DNA of glyphosate resistant rape was extracted by the method of CTAB, then reference gene BnACCg8, and three exoge⁃ nous genes of FMV 35S promoter, E9 3 terminator and CP4-EPSPS were amplified by PCR, and the amplified products were detected and analyzed on agar sugar gel electrophoresis. Results: There were three fragments which as same size as three exogenous genes of FMV 35S promoter, E9 3 terminator and the CP4