环介导等温扩增技术检测含有CaMV35S的转基因玉米

doi:10.7668/hbnxb.2011.04.002

华北农学报 ›› 2011, Vol. 26 ›› Issue (4): 8-14. doi: 10.7668/hbnxb.2011.04.002

所属专题： 玉米；生物技术；

环介导等温扩增技术检测含有CaMV35S的转基因玉米

陈金松^1,2, 黄丛林¹, 张秀海¹, 吴忠义¹

1. 北京市农林科学院北京农业生物技术研究中心, 北京 100097
2. 首都师范大学生命科学学院, 北京 100048

收稿日期:2010-10-29 出版日期:2011-08-28
通讯作者: 吴忠义(1969-)，男，福建泉州人，副研究员，主要从事植物逆境分子生物学研究。
作者简介:陈金松(1986-)，女，浙江温州人，在读硕士，主要从事生物化学与分子生物学研究。
基金资助:
国家转基因专项资助项目(2009ZX08003-009B);北京市科委项目(KJCX201102003)

Loop-mediated Isothermal Amplification for the Detection of CaMV35S Promoter in Genetically Modified Maize

CHEN Jin-song^1,2, HUANG Cong-lin¹, ZHANG Xiu-hai¹, WU Zhong-yi¹

1. Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;
2. College of Life Science, Capital Normal University, Beijing 100048, China

Received:2010-10-29 Published:2011-08-28

摘要/Abstract

摘要： DNA环介导等温扩增技术是一种特异、灵敏、快速的新型基因检测技术。针对玉米表达载体的花椰菜花叶病毒35 S启动子(CaMV35S)的6个区域设计4种特异引物,对LAMP反应的MgSO₄、dNTPs、Betaine、内引物、外引物各个成分进行了优化,此外还对LAMP和PCR两种不同方法的特异性进行了比较。建立转基因玉米花椰菜花叶病毒35 S启动子环介导等温扩增技术检测方法,LAMP与PCR相比具有更高的灵敏度。环介导等温扩增技术检测方法具有时间少,成本低,特异性高,检测方法多样等优势,为检测转基因玉米花椰菜花叶病毒35 S启动子提供了一种更加简便快速的方法。

关键词: 转基因玉米, 环介导等温扩增技术(LAMP), 检测方法

Abstract: The objective is to develop a loop-mediated isothermal amplification method for detection of CaMV-35S promoter in Genetically Modified maize.Loop-mediated isothermal amplification(LAMP),a novel nucleic acid amplification method,was developed for the rapid detection of Genetically Modified maize.Cauliflower mosaic virus 35S(CaMV35S) promoter gene was amplified by a set of four primers that recognize six distinct sequences of the target.The optimized conditions for LAMP were studied using different Mg²⁺,dNTPs,Betaine and primers concentrations.Furthermore,the sensitivity of LAMP was tested comparing with PCR.The LAMP method can detect CaMV35S promoter from genetically modified maize and their test results were consistent with the results of conventional PCR method.LAMP assay results were found to be more sensitive than the conventional PCR.The LAMP assay is an extremely rapid,highly sensitive,specific method,and will be an effective tool for rapid detection of Genetically Modified maize.

Key words: Genetically modified maize, Loop-mediated isothermal amplification(LAMP), Detection method

中图分类号:

S513.03

陈金松, 黄丛林, 张秀海, 吴忠义. 环介导等温扩增技术检测含有CaMV35S的转基因玉米[J]. 华北农学报, 2011, 26(4): 8-14. doi: 10.7668/hbnxb.2011.04.002.

CHEN Jin-song, HUANG Cong-lin, ZHANG Xiu-hai, WU Zhong-yi. Loop-mediated Isothermal Amplification for the Detection of CaMV35S Promoter in Genetically Modified Maize[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(4): 8-14. doi: 10.7668/hbnxb.2011.04.002.

http://www.hbnxb.net/CN/Y2011/V26/I4/8

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