转基因水稻Bt63的外源基因相对含量分析

Analysis of the Relative Content of Exogenous Gene in Transgenic Rice Bt63

为分析转基因Bt63稻米外源基因含量,利用新型、灵敏和高通量的实时荧光定量PCR技术,以RBE-4作为转基因水稻的内源参照基因,通过梯度稀释法,分别获得了Bt 63和RBE-4基因的Ct值与起始模板相关性的标准曲线,相关系数分别为0.996 6和0.995 4。通过转基因水稻外源基因(Bt63)和内标基因(RBE-4)起始模板数的比较,测定了外源基因在转基因水稻中的相对含量,这项研究将为建立转基因水稻抗虫能力评价体系和转基因稻谷的流通和监管提供技术支撑。

Abstract In order to analyze relative content of exogenous gene of transgenic rice Bt63, a new - type, sensitive and high throughput real -time fluorescence quantitative PCR technique was utilized in this experiment. /？BE-4 intransgenic rice was used as endogenous reference gene. The standard curves of the threshold cycle （Ct） relative to the log each initial template copy of Bt63 and RBE - 4 were obtained with a serial of dilutions and the correlation co- efficients were 0.996 6 and 0. 995 4, respectively. The relative content of exogenous gene in transgenic rice was ob- tained by comparing the initial template copy of exogenous gene Bt63 with that of endogenous reference gene RBE - 4. This result will set up the appraisement system of transgenic rice＇s insect resistance ability and a basis for detection and monitor of transzenic rice.