摘要
随着转基因技术的成熟,新型转基因产品相继问世,社会对其安全性的关注度也逐渐提高。而抗草丁膦(bialaphos resistance,Bar)基因作为转基因玉米中最常见的外源基因,常作为检测转基因的靶标元件。重组酶聚合酶扩增技术避开聚合酶链式技术反应时间长、程序繁琐和荧光定量、芯片等技术依赖高端设备,价格高昂等缺点。为简单、快速且不依赖于高端设备的转基因玉米检测新方法提供了新途径。本研究选取重组酶聚合酶扩增技术,设计并筛选最佳引物对,通过实验优化,最终得出最佳反应温度37℃及最佳时间为20 min的优化反应体系。经试验优化后的RPA扩增体系特异性良好,灵敏度可达fg级,为快速、恒温、现场检测提供了一种新手段。
With the maturity of transgenic technology,new transgenic products have come out one after another,and the attention of the society to its safety is gradually increasing.Bialaphos resistance(Bar),as the most common exogenous gene in transgenic maize,is often used as a target element in the detection of transgenic maize.Recombinant enzyme polymerase amplification(RPA)technology avoids the shortcomings of polymerase chain technology,such as long reaction time,complicated procedures,and fluorescence quantification and chip technology,which rely on high-end equipment and high price.It provides a new way for a simple,rapid and independent detection method of transgenic maize.In this paper,the recombinant enzyme polymerase amplification technology was selected to design and screen the best primer pairs.Through experimental optimization,the optimal reaction system with the best reaction temperature of 37℃and the best time of 20 min was finally obtained.The optimized RPA amplification system has good specificity and sensitivity up to fg,which provides a new method for rapid,constant temperature and in situ detection.
作者
段瑞杰
焦晓歌
黄勇
周宇航
张辰宇
徐亚维
Duan Ruijie;Jiao Xiaoge;Huang Yong;Zhou Yuhang;Zhang Chenyu;Xu Yawei(School of Biological and Pharmaceutical Engineering,Jilin Agricutural Science and Technology University,Jilin,132101)
出处
《分子植物育种》
CAS
北大核心
2022年第22期7494-7500,共7页
Molecular Plant Breeding
基金
吉林省大学生科技创新项目(201911493014)
吉林省科技厅项目(20200403053SF)共同资助。
关键词
转基因
草丁膦
重组酶聚合酶扩增
特异性
灵敏度
Transgenic
Bialaphos resistance
Recombinant enzyme polymerase amplification
Specificity
Sensitivity
