生物
环介导等温扩增
可选择标记
基因
聚合酶链反应
转基因作物
分子生物学
转基因
转基因生物
作者
Jinsong Chen,Conglin Huang,Xiuhai Zhang,Rong Yu,Zhongyi Wu
出处
期刊:African Journal of Biotechnology
[Academic Journals]
日期:2011-11-28
卷期号:10 (75): 17055-17061
被引量:10
摘要
Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method that reagents react to under isothermal conditions with high specificity, efficiency and rapidity. We used LAMP for detection of herbicide-resistant maize, which is a widespread genetically modified crop. Pat gene, a widely used selectable marker, was amplified by a set of four special primers that recognized six distinct sequences of the target. We studied the optimized conditions for LAMP using different Mg 2+ , dNTPs, betaine and primers concentrations. Furthermore, we tested the sensitivity and specificity of LAMP as compared to PCR. This work shows that the LAMP method can detect a specific pat gene from herbicide-resistant maize and their test results are consistent with the results of the conventional PCR methods. However, LAMP assay results were found to be more sensitive than the conventional PCR. Key words: Herbicide-resistant maize, loop-mediated isothermal amplification (LAMP), transgenic maize detection, pat gene.
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