SYBR~ Green I实时荧光PCR鉴定检测转基因油菜RT73品系

Identification for Genetically Modified Canola RT73 Line with SYBR~ Green I Real Time PCR

根据转基因油菜RT73品系转入的外源基因质粒图谱,设计品系鉴定的特异性引物,进行SYBR!Green I实时PCR检测,建立转基因油菜品系鉴定的SYBR Green I实时PCR方法。同时针对油菜FatA基因设计出内源基因。结果表明:应用实时PCR技术,利用SYBR Green I染料能选择性结合双链DNA的特点,可检测到RT73品系特异性靶序列扩增所产生的荧光信号,且通过溶解曲线确定其熔点值为80.2(±0.5)。SYBR Green I实时PCR能通过溶解曲线有效地区分特异性产物、非特异性产物以及引物二聚体,是鉴定转基因油菜RT73品系的新方法。

To identify genetically modified （GM） canola RT73 line, a SYBR Green I real-time PCR assay was performed in this study. Specific primers for inserted genes in the RT73 were used to conduct the SYBR Green I real-time PCR assays. The SYBR Green I real-time PCR method was established to detect and identify GM canola lines. The endogenesis FatA gene of canola could be designed to control the testing process and avoid the fake negative result. The results showed that the SYBR Green I could selectively combine double chain DNA, identify RT73 canola, and confirm melting point of 80.2（＋0.5） by melting curve, while other GM and NO-GM canola didn＇t be detected. The SYBR Green I real-time PCR could be a new method for detecting genetically modified （GM） canola RT73 line.