SYBR Green实时荧光定量PCR检测大豆转基因成分

doi:10.7506/spkx1002-6630-200908035

食品科学 ›› 2009, Vol. 30 ›› Issue (8): 171-176.doi: 10.7506/spkx1002-6630-200908035

SYBR Green实时荧光定量PCR检测大豆转基因成分

王小花¹，李建祥¹，王国卿²，李新莉¹，邵景东³，傅春玲^1,*

1.苏州大学放射医学与公共卫生学院 2.苏州大学基础医学与生物科学学院 3.张家港出入境检验检疫局

收稿日期:2008-06-08 修回日期:2008-09-04 出版日期:2009-04-15 发布日期:2010-12-29
通讯作者: 傅春玲1,* E-mail:fuchunling@suda.edu.cn
基金资助:
江苏省高校自然科学基金项目(06KJD330159)

Detection of Transgenic Components in Soybean Products by SYBR Green Real-Time PCR

WANG Xiao-hua1 LI Jian-xiang1 WANG Guo-qing2 LI Xin-li1 SHAO Jing-dong3 FU Chun-ling1,*

(1. College of Radiation Medicine and Public Health, Soochow University, Suzhou 215123, China
2. College of Preclinical Medicine and Life Science, Soochow University, Suzhou 215123, China
3. Zhangjiagang Entry-Exit Inspection and Quarantine Bureau, Zhangjiagang 215633, China)

Received:2008-06-08 Revised:2008-09-04 Online:2009-04-15 Published:2010-12-29
Contact: FU Chun-ling1,* E-mail:fuchunling@suda.edu.cn

摘要/Abstract

摘要：

采用SYBR Green实时荧光定量PCR技术，建立了食品中大豆转基因成分的定量检测方法。通过设计特异引物，扩增内源参照基因lectin和转基因靶基因CP4EPSPS，建立两种基因的拷贝数-CT标准曲线，根据标准曲线方程计算样品中的转基因含量，并且通过熔解曲线分析扩增反应特异性。结果表明，lectin和CP4EPSPS基因标准曲线线性关系好，R2值分别为0.9984和0.9953，方法的回收率为95%～110%，检测限为0.01%。本检测方法具有快速、灵敏、准确、特异、高通量等优点，可以作为食品中大豆转基因成分的定量检测方法。

关键词: 实时定量PCR, SYBR GreenⅠ荧光染料, 转基因大豆

Abstract:

A quantitative method to detect the transgenic component in soybean by using real-time PCR technique based on fluorescence dye SYBR Green was investigated in this study. The endogenous lectin gene and transgenic target CP4EPSPS gene were amplified through the design of specific primers, and the transgenic content was then calculated according to the standard curve equation. Meanwhile the specificity of PCR amplification was analyzed with corresponding melting curves .The results showed that the standard curves of lectin and CP4EPSPS genes have good linear relationship, and their R2 values are 0.9984 and 0.9953, respectively. Moreover, the recovery rate of the method is between 95% and 110% and the detection limit is 0.01%. In conclusion, this method is fast, sensitive, simple, accurate, specific and of high throughput, and can be used to quantificationally detect transgenic components in soybean products.

Key words: real-time quantitative PCR, fluorescence dye SYBR Green I, transgenic soybean

中图分类号:

Q503

王小花1，李建祥1，王国卿2，李新莉1，邵景东3，傅春玲1,*. SYBR Green实时荧光定量PCR检测大豆转基因成分[J]. 食品科学, 2009, 30(8): 171-176.

WANG Xiao-hua1 LI Jian-xiang1 WANG Guo-qing2 LI Xin-li1 SHAO Jing-dong3 FU Chun-ling1,*. Detection of Transgenic Components in Soybean Products by SYBR Green Real-Time PCR[J]. FOOD SCIENCE, 2009, 30(8): 171-176.

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SYBR Green实时荧光定量PCR检测大豆转基因成分

Detection of Transgenic Components in Soybean Products by SYBR Green Real-Time PCR

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