转基因大豆论文转基因食物论文

基因组学与应用生物学，2010年，第29卷，第6期，第1177－1183页

Genomics and Applied Biology,2010,Vol.29,No.6,1177－1183

技术改进

Upgrated Technology

转基因大豆GTS40－3－2转化事件特异性PCR检测

王恒波1,2陈平华1,2郭晋隆1陈如凯1*许莉萍1

1福建农林大学农业部甘蔗遗传改良重点开放实验室,福州,350002;2农业部甘蔗及制品监督检验测试中心转基因生物产品检测室,福州, 350002

*通讯作者,phcemail@

摘要GTS40－3－2是抗草甘膦转基因大豆，为建立GTS40－3－2大豆转化体特异性PCR检测方法，本研究以GTS40－3－2标准品为实验材料，根据已公布转基因大豆GTS40－3－2基因与大豆基因组连接序列信息，利用Primer5.0软件设计了5对品系特异性引物，对每对引物进行了退火温度、特异性及扩增效率的PCR检测，结果显示，5对特异性引物均能够从GTS40－3－2中扩增出大小约279bp、238bp、470bp、490bp和257bp 的预期产物，可用于特异性检测转基因大豆GTS40－3－2转化事件。以转基因大豆GTS40－3－2含量为5%、2%、1%、0.5%和0.1%的标准品进行PCR灵敏度检测，结果表明5对引物的检测灵敏度均能达到0.1%。通过荧光定量PCR对5对特异性引物的Ct值与溶解曲线比较，最后选择出RRS2引物对为转基因大豆GTS40－3－2品系特异性检测的最适引物。本文结果将为我国未来转基因生物产品成分检测提供科学合理的实验参考。

关键词转基因大豆,GTS40－3－2,转化事件,特异性PCR检测

Specific PCR Validation of Transformation Event for Transgenic Soybean GTS40－3－2

Wang Hengbo1,2Chen Pinghua1,2Guo Jinlong1Chen Rukai1*Xu Liping1

1Key Laboratory of Sugarcane Genetic Improvement,Ministry of Agriculture,Fujian Agriculture and Forestry University,Fuzhou,350002;2Quality Supervision,Inspection and Testing Center for Sugarcane and Derived Products of Ministry of Agriculture,Fuzhou,350002

*Corresponding author,phcemail@

DOI:10.3969/gab.029.001177

Abstract GTS40－3－2is a glyphosate-tolerant transgenic soybean.In order to establish a specific PCR validation method for transgenic soybean GTS40－3－2,in this research,we employed GTS40－3－2standard sample as the ex-periment material,and designed five specific PCR primers based on the junction sequence published between transgenic soybean gene GTS40－3－2and soybean genome by using Primer5.0software to PCR validation the Tm value,specific and the amplification efficiencies from each primer.The results showed that the products size of five primers was in accordance with expected one which was279bp,238bp,470bp,490bp and257bp respec-tively from the standard reference GTS40－3－2and they can be used for specific validation of transformation event in transgenic soybean GTS40－3－2.Further,we utilized the content of5%,2%,1%,0.5%and0.1%transgenic soy-bean GTS40－3－2as the sample to carry out PCR sensitivity validation,the results demonstrated the sensitivity of the five primers could reach0.1%.Finally,we selected out RRS2primer was the best primer for specific validation of transformation event in transgenic soybean GTS40－3－2by using fluorescent quantitative PCR in comparison with Ct values and the melting curves among five specific Primers.The results of this paper would provide a scien-

基金项目：本研究由现代农业产业技术体系建设专项资金(nycytx－24)、国家948项目(2006－G37,2010－S19)、国家基金项目(310 70330)、国家“863”计划项目(2007AA100701)、福建省科技计划重点项目(2007I0036)、福建省自然科学基金项目(2009J05050)和福建科技项目备案(F2007AA100701)共同资助