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若干定性PCR方法部颁标准在检测转基因混合样品中的应用分析
引用本文:李美英,易小平,杨小亮,肖苏生,郭安平.若干定性PCR方法部颁标准在检测转基因混合样品中的应用分析[J].热带作物学报,2013,34(10):1955-1960.
作者姓名:李美英  易小平  杨小亮  肖苏生  郭安平
作者单位:1 中国热带农业科学院热带生物技术研究所农业部热带作物生物技术与遗传资源利用重点实验室 2 农业部转基因植物及植物用微生物环境安全监督检验测试中心;1 中国热带农业科学院热带生物技术研究所农业部热带作物生物技术与遗传资源利用重点实验室 2 农业部转基因植物及植物用微生物环境安全监督检验测试中心;1 中国热带农业科学院热带生物技术研究所农业部热带作物生物技术与遗传资源利用重点实验室 2 农业部转基因植物及植物用微生物环境安全监督检验测试中心;1 中国热带农业科学院热带生物技术研究所农业部热带作物生物技术与遗传资源利用重点实验室 2 农业部转基因植物及植物用微生物环境安全监督检验测试中心;1 中国热带农业科学院热带生物技术研究所农业部热带作物生物技术与遗传资源利用重点实验室 2 农业部转基因植物及植物用微生物环境安全监督检验测试中心
基金项目:转基因作物南繁区环境安全监测与控制(No. 2013ZX08012-004);现代农业人才支撑计划;中央级公益性科研院所基本科研业务费资金(No. ITBBZX0833,No. ITBB110603)。
摘    要:以若干定性PCR方法部颁标准对含有0.5%的4份不同转基因混合样品进行检测,先以通用元件标准中的CaMV35s启动子、NOS终止子对混合样品进行初步定性PCR筛选。结果表明,4份样品中都含有转基因成分。Bt基因特异性标准检测表明,3#和4#样品含有转基因抗虫水稻成分。构建特异性标准PCR检测表明,2#、3#和4#样品含有转基因GTS-40-3-2大豆成分。以MON810、Bt176、NK603转化体事件标准进行品系特异性PCR检测,结果证实:1#和4#样品中含有Bt176转基因玉米成分;3#样品中含有Mon810转基因玉米成分;4份样品中均不含NK603转基因玉米成分。说明农业部颁布的定性PCR方法标准能满足于对多种转基因混合样品的检测,且检测结果准确,可靠。

关 键 词:转基因混合样品,定性PCR,检测

Application of Several Qualitative Polymerase Chain Reaction (PCR)Detection Standards Issued by Ministry of Agriculture in the Detection of Transgenic Mixture Samples
LI Meiying,YI Xiaoping,YANG Xiaoliang,XIAO Susheng and GUO Anpin.Application of Several Qualitative Polymerase Chain Reaction (PCR)Detection Standards Issued by Ministry of Agriculture in the Detection of Transgenic Mixture Samples[J].Chinese Journal of Tropical Crops,2013,34(10):1955-1960.
Authors:LI Meiying  YI Xiaoping  YANG Xiaoliang  XIAO Susheng and GUO Anpin
Institution:1 Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agriculture Sciences / Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture 2 Environmental Safety Supervision and Inspection Center;1 Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agriculture Sciences / Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture 2 Environmental Safety Supervision and Inspection Center;1 Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agriculture Sciences / Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture 2 Environmental Safety Supervision and Inspection Center;1 Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agriculture Sciences / Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture 2 Environmental Safety Supervision and Inspection Center;1 Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agriculture Sciences / Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture 2 Environmental Safety Supervision and Inspection Center
Abstract:Mixed samples containing 0.5% of transgenic maize, transgenic rice, and transgenic soybean were used to verify several qualitative polymerase chain reactions(PCR)analysis standards issued by Ministry of Agriculture in the detection of transgenic mixture products. Firstly, CaMV35s promoter and NOS terminator PCR detection standard were used to screen the mixed samples and indicated the four mixed samples were transgenic products initially. The specificity of BT PCR detection standard indicated 3# and 4# samples contained transgenic BT rice, construction specific PCR detection standard indicated 2#, 3#, and 4# samples contained transgenic GTS-40-3-2 soybeans. Transformation body events PCR detection standard suggested 1# and 4# samples contained transgenic Bt176 maize, and 3# samples contained Mon810 transgenic maize. The results suggested the general qualitative polymerase chain reaction detection standards issued by Ministry of Agriculture are sufficient and reliable to detect mixted genetically modified products.
Keywords:Transgenic mixture products  Qualitative polymerase chain reaction  Detection
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