|Table of Contents|

Rapid Detection of Five Genetically Modified Soybean Lines by Multiplex PCR Method(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2016年06期
Page:
34-39
Research Field:
Publishing date:

Info

Title:
Rapid Detection of Five Genetically Modified Soybean Lines by Multiplex PCR Method
Author(s):
DONG Li-ming; LI Cong-cong; XING Zhen-juan; SHAO Gai-ge; XIA Wei; YAN Wei; LI Fei-wu
（Institute of Agricultural Quality Standard and Testing Technology，Jilin Academy of Agricultural Sciences，Changchun 130033，China）
Keywords:
Genetically modified soybean; Multiplex PCR; Event-specific detection
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2016.06.1002
Abstract:
The soybean lines 305423，MON89788，CV127 and GTS40-3-2 are the most widely commercialized genetically modified soybeans and account for more than 80% of the world total transgenic soybean planting acreage. In this study, the specific primers were designed according to the border sequences of five GM soybean events.By verifying the applicability，specificity and sensitivity of the primers the multiplex PCR conditions，including the amount of primers and the annealing temperature，were optimized. Then a multiplex PCR detection system which can be used to amplify soybean -lectin- gene and introduced genes in five GM soybean events simultaneously was developed. The results showed that the primers applied in this method had good specificity，no cross-amplification and non-specific amplification. The detection limit of this method could reach 0.1%.This method could be used as an auxiliary means to detect the genetically modified ingredients rapidly.

References:

［1］James C. 2015年全球生物技术/转基因作物商品化发展态势［J］. 中国生物工程杂志，2016，36(4)：12-17.（James C. Global development trend of commercialized biotech/GM crops in 2015［J］.China Biotechnology，2016，36(4)：12-17.）

［2］罗阿东，焦彦朝，曹云恒等. 转基因大豆检测技术研究进展［J］. 南方农业学报，2012，43(3)：290-293.（Luo A D，Jiao Y C，Cao Y H，et al. Progresses in detection technologies for transgenic soybean ［J］. Journal of Southern Agriculture，2012，43(3)：290-293.）
［3］张大兵，郭金超. 转基因生物及其产品检测技术和标准化［J］. 生命科学，2011，23(2)：195-204.（Zhang D B，Guo J C.The development and standardization of testing approaches for genetically modified organisms and their derived products ［J］. Chinese Bulletin of Life Sciences，2011，23(2)：195-204.）
［4］Shao N，Jiang S M，Zhang M，et al. MACRO：A combined microchip-PCR and microarray system for highthroughput monitoring of genetically modified organisms ［J］. Analytical Chemistry，2014，86(2)： 1269-1276.
［5］Heide B R，Heir E，Holck A. Detection of eight GMO maize events by qualitative,multiplex PCR and fluorescence capillary gel electrophoresis ［J］ European Food Research Technology，2008，227：527-535.
［6］Xu W T，Zhai Z F，Huang K L，et al. A novel universal Primer-Multiplex-PCR method with sequencing gel electrophoresis analysis ［J］. PLoS One，2012，7(1)：e22900.
［7］傅凯，黄文胜，邓婷婷等. 多重PCR-液相芯片技术检测13个品系转基因玉米［J］. 中国食品学报，2015，15(1)：188-197（Fu K，Huang W S，Deng T T，et al. Multiplex PCR assay and liquid bead array for detection of 13 lines genetically modified maize ［J］. Journal of Chinese Institute of Food Science and Technology，2015，15(1)：188-197.）
［8］Li F W，Yan W，Long L K，et al. Screening detection of Bt genes in genetically modified crops using a multiplex PCR assay ［J］. Modern Food Science & Technology，2014，30(5)：262-266.
［9］NY/T 675-2003，转基因植物及其产品检测—大豆定性PCR方法［S］. 北京：中国标准出版社，2003.（Agricultural industry standard of the People′s Republic of China NY/T675-2003. Detection of genetically modified plant organisms and derived products-qualitative PCR methods for soybeans ［S］. Beijing：Standards Press of China，2003）
［10］农业部1782号公告1-2012，转基因植物及其产品成分检测—耐除草剂大豆356043及其衍生品种定性PCR方法［S］. 北京：中国标准出版社，2012（Announcement No. 1782-1-2012 of the Ministry of Agriculture Detection of genelically modified plants and derived products-qualitative PCR method for herbicide-tolerant soybean 356043 and its derivates ［S］. Beijing:Standards Press of China，2012.）
?［11］Liu J，Guo J C，Zhang H B，et al. Development and in-house validation of the event-specific polymerase chain reaction detection methods for genetically modified soybean MON89788 based on the cloned integration flanking sequence ［J］. Journal of Agriculture and Food Chemistry，2009，57:10524-10530.
［12］农业部1782号公告5-2012，转基因植物及其产品成分检测—耐除草剂大豆CV127及其衍生品种定性PCR方法［S］. 北京：中国标准出版社，2012.（Announcement No. 1782-5-2012 of the Ministry of Agriculture Detection of genetically modified plants and derived products-qualitative PCR method for herbicidetolerant soybean CV127 and its derivates ［S］. Beijing：Standards Press of China，2012.）
［13］农业部1861号公告2-2012，转基因植物及其产品成分检测—耐除草剂大豆GTS40-3-2及其衍生品种定性PCR方法［S］. 北京：中国标准出版社，2012（Announcement No. 1861-2-2012 of the Ministry of Agriculture Detection of genelically modified plants and derived products-Qualitative PCR method for herbicide-tolerant soybean GTS 40-3-2 and its derivates ［S］. Beijing：Standards Press of China，2012.）

Memo

Memo:
-
Last Update: 2016-12-09