荧光PCR方法定性和定量检测BT63转基因大米

QUALITATIVE AND QUANTITATIVE DETERMINATION OF GENETICALLY MODIFIED BT-RICE

采用实时荧光定量PCR技术,建立定性(量)鉴定转BT基因稻米成分的方法。根据稻米内源蔗糖磷酸合成酶(SPS)和结构特异性BT基因序列的特点设计引物和探针,可特异性地对转BT基因的稻米进行品种鉴定。把100%转BT基因稻米提取得到的DNA进行梯度稀释,作为标准DNA构建荧光定量标准曲线。检测限为0.5%,在0.5%水平上检测误差为20%。

Fluorescent qualitative and quantitative method were firstly developed for the detection of Genetically Modified BT-rice. Based on endogenous control gene （sps） and construct-specific gene （BT） sequence, we designed target-specific primers and fluorescent probes for the identification of Genetically Modified BT-rice. DNA was extracted and diluted which as standard DNA to gain standard curve. PCR was performed on genomic DNAs extracted from BT-rice,and other GM events. As reference DNAs, 100 % DNA samples was serial diluted （lO-fold）. Limit of detection of the method was 0.5 %. At the 0.5 % level, the bias （mean vs true value） for BT-rice was 20.0 %.