[目的]探讨SYBR Green实时定量PCR技术应用于检测转基因植物外源基因拷贝数的可行性.[方法]使用SYBR Green实时定量PCR技术,以转CYCD3;1的拟南芥为材料,通过CYCD3;1基因与单拷贝内参基因以及转基因株系中的CYCD3;1基因与野生型植株的比较来确定外源基因的拷贝数.[结果]该法计算所得外源基因拷贝数与传统高准确性的Southern blot法结果相符.[结论]使用该法检测外源基因拷贝数简单易行,可操作性强.%[Objective] To explore the feasibility of using SYBR Green real-time quantitative PCR technique to estimate the transgene copy number of a transgenic plant. [ Method] Using SYBR Green real-time quantitative PCR technique, we have determined the copy numbers of the exogenous CYCD3 ; 1 in transgenic plants by comparing an endogenous single copy reference gene with CYCD3; 1 copy number in transgenic plants, meanwhile comparing CYCD3;1 copy numbers between wild plant and transgenic plants. [Result]The exogenous CYCD3;1 copy num bers calculated by this method is identical with results of traditional Southern blot analysis which is highly accurate. [ Conclusion ] This method is simple, effective and safe for estimating transgene copy numbers.
展开▼
