Development and application of loop-mediated isothermal amplification for detection of the cry1Ac gene in transgenic insect-resistant rice

To develop a convenient and rapid assay method for identification of the modified cry1Ac gene in the transgenic insect-resistant rice,a loop-mediated isothermal amplification(LAMP) analysis method was established.The target DNA fragment of the modified cry1Ac gene in transgenic insect-resistant rice was successfully amplified in a LAMP reaction system with Bst DNA polymerase and a set of four special primers that recognized six distinct sequences of the target gene after incubation at 65℃for 60min.The sensitivity of the LAMP cn.method was 10times of that of the conventional PCR method.It could detect the modified cry1Ac gene in a sample consisting of the transgenic insect-resistant rice and non-transgenic rice at a ratio as low as 0.01%.Two genetically modified crops(GMCs) and five non-GMCs were identified using the LAMP method and the results demonstrated that it could detect the modified cry1Ac gene in GMCs with high specificity.Thus the LAMP method will be potentially an effective tool for identification of GMCs by the rapid detection of the modified cry1Ac gene in transgenic insect-resistant crops.