|本期目录/Table of Contents|

[1]王恒波,陈如凯,陈平华.抗除草剂转基因作物实时荧光定量PCR检测[J].应用与环境生物学报,2009,15(06):866-870.[doi:10.3724/SP.J.1145.2009.00866]
 WANG Hengbo,CHEN Rukai,CHEN Pinghua.Detection of Genetically Modified Herbicide–tolerant Crops by Real-time Fluorescent Quantitative PCR Assay[J].Chinese Journal of Applied & Environmental Biology,2009,15(06):866-870.[doi:10.3724/SP.J.1145.2009.00866]
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抗除草剂转基因作物实时荧光定量PCR检测()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
15卷
期数:
2009年06期
页码:
866-870
栏目:
技术与方法
出版日期:
2009-12-25

文章信息/Info

Title:
Detection of Genetically Modified Herbicide–tolerant Crops by Real-time Fluorescent Quantitative PCR Assay
作者:
王恒波陈如凯陈平华
(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)
Author(s):
WANG Hengbo CHEN Rukai CHEN Pinghua
(Key Laboratory of Genetic Improvement for Sugarcane, Ministry of Agriculture, Sugarcane Research Institute of
Fujian Agriculture and Forestry University, Fuzhou 350002, China )
关键词:
抗除草剂转基因作物SYBR Green I荧光染料实时荧光定量PCR溶解曲线检测
Keywords:
genetically modified herbicide-tolerant crop SYBR Green I dye real-time fluorescent quantitative PCR melting curve detection
分类号:
Q943.2
DOI:
10.3724/SP.J.1145.2009.00866
文献标志码:
A
摘要:
建立了一种以SYBR Green I为结合染料、快速准确检测转抗除草剂基因成分的实时荧光定量PCR方法. 以转基因大豆与转基因玉米标准品为材料,通过使用特异性引物和SYBR Green I结合染料实时荧光定量PCR技术,对转基因农作物中外源抗除草剂基因进行了定量检测,绘制了两种基因扩增的标准曲线图,根据标准曲线方程计算外源基因含量;并作了溶解曲线、检测方法检测灵敏度和精密度的分析. 研究发现,两者标准曲线方程线性关系良好,R2值分别达到0.993 9与0.992 4. 通过已知标准品进行验证,实测值与真值接近,与实际含量的相对偏差是6.52%和7.90%. 结果表明,SYBR Green I结合染料法完全可以用于转基因农作物定量PCR检测. 图5 表2 参11
Abstract:
A rapid and sensitive SYBR Green I-based real-time PCR method was developed for quantitative detection of CP4-EPSPS and PAT genes from the genetically modified (GM) herbicide-tolerant crops. The target genes from transgenic soybean and maize references were amplified to make standard curves. The transgenic ratios were then calculated according to the standard Ct-copies linear graphs of these two genes. The reproducibility and melting curves of the genes were also analyzed. The results showed that the standard equations of CP4-EPSPS and PAT genes had higher R2 values of 0.993 9 and 0.992 4, respectively. The difference in transgenic ratios between the known standards and measured crops was only 6.52%~7.90%. These results suggest that the real-time PCR assay with SYBR Green I dye is suitable for detecting the ratios of GM crops and their derivates. Fig 5, Tab 2, Ref 11

参考文献/References:

1 Zhang LG (张立国), Zhang J (张琚). Introduced the quantitative real-time PCR technology. Biotechnology (生物技术), 2003, 13 (2): 39~40
2 Zhu YZ (朱元招), Yin JD (尹靖东), Li DF (李德发), Wang FL (王凤来). Quantitative PCR method for detecting glyphosate tolerant gene transfer in soybeans. J China Agric Univ (中国农业大学学报), 2005, 10 (3): 25~29
3 Knut G, Berdal K, Holst-Jensen A. Roundup Ready soybean event-specific real-time quantitative PCR assay and estimation of the practical detection and quantification limits in GMO analyses. Eur Food Res Technol, 2001, 2 (13): 432~438
4 Vailingom M, Pijnenburg H, Gendre F, Brignon P. Real-time quantitative PCR detection of genetically modified maximizer maize and Roundup Ready soybean in some representative foods. J Agric Food Chem, 1999, 47: 5261~5266
5 Chen Y (陈颖), Xu BL (徐宝良), Zhu N (朱宁), Ge YQ (葛毅强), Wang SG (王曙光). Real-time quantitative PCR detection of genetically modified maximizer maize and yieldgard maize. Acta Agron Sin (作物学报), 2004, 6 (30): 602~607
6 Knut GB, Arne HJ. Roundup Ready soybean event-specific real-time quantitative PCR assay and estimation of the practical detection and quantification limits in GMO analysis. Eur Food Res Technol, 2001, 2 (13): 432~438
7 Chen JQ (陈建魁). Quantitative PCR technology. J Clin Lab (临床检验杂志), 1997, 15 (2): 121~123
8 Pan LW (潘良文), Chen JH (陈家华), Luo D (罗达), Yu DY (喻德跃), Feng XZ (冯献忠). Quantitative detection of genetically modified Bt176 maize in maize powder. J Plant Physiol & Mol Biol (植物生理与分子生物学学报), 2002, 28 (6): 463~467
9 Tan W (谭文), Cao JJ (曹际娟), Zhu SF (朱水芳). Quantitative identified detection of the genetically modified maize Bt11 components in processed products. Biotechnol Bull (生物技术通讯), 2003 (6): 46~50
10 Ririe KM, Rasmussen RP, Wittwer CT. Product differentiation by analysis of DNA melting curves during the polymerase chain reaction. Anal Biochem,1997, 245: 154~160
11 Mackay IM, Arden KE, Nitsche A. Real-time PCR in virology. Nucleic Acids Res, 2002, 30 (6): 1292~1305

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备注/Memo

备注/Memo:
国家“863”计划项目(No. 2007AA100701)和福建省科技计划重点项目(No. 2007I0036)资助 Support by the National “863” Project of China (No. 2007AA100701) and the Major Research Projects of the Department of Science & Technology of Fujian, China (No. 2007I0036)
更新日期/Last Update: 2009-12-24