|本期目录/Table of Contents|

[1]王嘉钰 董娟** 唐卓**.基于环介导等温扩增法可视化检测玉米转基因成分*[J].应用与环境生物学报,2019,25(06):1-9.[doi:10.19675/j.cnki.1006-687x.2019.04020]
 WANG Jiayu,DONG Juan** & TANG Zhuo**.Colorimetric detection of genetically modified organisms (GMO s) based on loop-mediated isoth ermal amplification (LAMP)*[J].Chinese Journal of Applied & Environmental Biology,2019,25(06):1-9.[doi:10.19675/j.cnki.1006-687x.2019.04020]
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基于环介导等温扩增法可视化检测玉米转基因成分*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年06期
页码:
1-9
栏目:
技术与方法
出版日期:
2019-12-30

文章信息/Info

Title:
Colorimetric detection of genetically modified organisms (GMO s) based on loop-mediated isoth ermal amplification (LAMP)*
作者:
王嘉钰12 董娟1** 唐卓1**
1中国科学院成都生物研究所 成都 610041
2中国科学院大学 北京 100049
Author(s):
WANG Jiayu1 2 DONG Juan1** & TANG Zhuo1**
1 Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China
2 University of Chinese Academy of Sciences, Beijing 100049, China
关键词:
环介导等温扩增转基因基因特异性可视化检测现场检测
Keywords:
loop-mediated isothermal amplification (LAMP) genetically modified (GM) gene-specific visualdetection on-site detection
DOI:
10.19675/j.cnki.1006-687x.2019.04020
摘要:
快速准确的转基因检测技术是严格监管转基因作物及其产品的重要支撑;不同转基因作物由于导入不同外源目的基因从而表现出不同性状,因此外源目的基因序列可作为基于核酸检测技术进行转基因筛选的标志物。基于环介导等温扩增(Loop-mediated isothermal amplification,LAMP)技术,以中性红染料作为比色试剂,建立玉米内源基因和玉米转基因成分的基因特异性的可视化快速检测方法。针对玉米内源基因IVR基因,抗虫外源bar、pat基因,和Bt176中的抗除草剂外源cry1Ab基因分别设计了LAMP引物。结果表明,在Real-time LAMP体系下,能够实现对玉米内源IVR基因、转基因玉米品系Bt176中的bar基因和转基因玉米品系TC1507和59122中的pat基因的检测。对转基因玉米品系Bt176中的cry1Ab基因设计的引物能特异性扩增Bt176,而对Bt11和MON810两种转基因玉米品系无效。通过优化Tris-HCl和中性红浓度,在5 mmol/L Tris-HCl和100 μmol/L 中性红浓度条件下建立了可视化检测体系,反应1 h即可分别实现对玉米内源基因IVR基因,抗虫外源bar、pat基因,和抗除草剂外源cry1Ab基因的检测。中性红可视化检测结果与Real-time LAMP荧光扩增曲线结果一致,体现了该可视化检测方法具有较高的准确性。由此,本研究建立了一种对玉米转基因相关成分的可视化检测体系,该方法既快速又简便且尤其适用于现场检测。(图6 表1 参17)
Abstract:
With the rapid development and application of genetically modified (GM) technology, its imperative to strictly supervise GM crops and their products . The rapid and accurate GMOs testing technology is an important support to complete this work. GM crops often contain different exogenous gene for various transgenic resistance. Therefore, these exogenous genes can be used as markers for screening and identifying the transgenic components based on nucleic acid detection technology. Here, the study was based on Loop-mediated isothermal amplification (LAMP) technology coupled with neutral red, an indicator dye, to establish colorimetric detection method for maize endogenous gene and common-used exogenous gene. Specific primers for endogenous gene IVR of maize and exogenous gene bar, pat, cry1 Ab were designed ba sed on LAMP, respectively. The results show that specific detection of IVR gene, bar gene in GM maize Bt176, pat gene in GM maize TC1507 and 59122 can be accomplished based on real-time LAMP fluorescence repor ting system. The here designed cry1Ab gene LAMP primers can specifically amplified GM maize Bt176, but invalid to GM maize Bt11 and MON810. The visual detection method was developed under the concentration of 5 mmol/L Tris-HCl and 100 μmol/L neutral red dye. The method was capable of detecting IVR gene and exogenous gene bar, pat, cry1Ab genes within 1 h . Based on our results, the visual results were consistent to r eal-time fluorescence LAMP results, indicating that our visual detection method is accurate and reliable . In conclusion, the here established visual LAMP method is capable of detecting GM maize. This method is rapid, simple, and shows the great potential of on-site detection.

相似文献/References:

[1]赵高梅,龙海,邓光兵,等.青稞B-hordein基因表达载体构建及对小麦的遗传转化[J].应用与环境生物学报,2011,17(03):364.[doi:10.3724/SP.J.1145.2011.00364]
 ZHAO Gaomei,LONG Hai,DENG Guangbing,et al.Vector Construction and Genetic Transformation of B-hordein Gene from Hulless Barley to Wheat[J].Chinese Journal of Applied & Environmental Biology,2011,17(06):364.[doi:10.3724/SP.J.1145.2011.00364]
[2]丁盛陈刚毅李美王嘉钰陈蓉邹家尉杜凤董娟黄鑫崔欣唐卓**.基于LAMP的病原菌一步等温可视化检测*[J].应用与环境生物学报,2019,25(05):1.[doi:10.19675/j.cnki.1006-687x.2019.01041]
 DING Sheng,CHEN Gangyi,et al.One-step isothermal visual detection of pathogens based on LAMP*[J].Chinese Journal of Applied & Environmental Biology,2019,25(06):1.[doi:10.19675/j.cnki.1006-687x.2019.01041]

备注/Memo

备注/Memo:
收稿日期 Received: 2019-04-09 接受日期 Accepted: 2019-04-30
四川省青年科技创新研究团队项目(2017TD0021)和成都市科技项目(2016-HM01-00371-SF)资助
**通讯作者 Corresponding author (E-mail: tangzhuo@cib.ac.cn, dongjuan@cib.ac.cn)
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更新日期/Last Update: 2019-04-30