应用hiTAIL-PCR扩增转基因木瓜的侧翼序列

High-efficiency thermal asymmetric interlaced PCR for amplification of flanking sequence of transgenic papaya lines

品系特异性检测是实施转基因产品标识管理的技术保障,而获取转基因品系的侧翼序列是建立品系特异性检测方法的前提。本研究应用hi TAIL-PCR扩增两个抗病毒转基因木瓜品系的RB区侧翼序列。结果表明,海南木瓜样品为转基因木瓜品系18-2-4,送检木瓜样品为转基因木瓜品系16-0-1。与传统TAIL-PCR方法相比,hi TAIL-PCR扩增的非目标片段和小片段少,获得目标片段的成功率高,是快速扩增已知序列邻近的侧翼序列的好方法。

Event-specific detection is the technical guarantee for implementing labeling regulations of genetically modified organism,and the acquisition of the flanking sequence is the prerequisite for developing the event-specific detection method. In this study,we amplified RB flanking sequences of two antiviral transgenic papaya lines by means of high-efficiency thermal asymmetric interlaced PCR（hi TAIL-PCR）.The results showed that the papaya sample from Hainan was the transgenic line 18-2-4,and submitted samples were all the transgenic line 16-0-1. Compared with the traditional TAIL-PCR method,the non-target fragments and small fragments by hi TAIL-PCR are few,and the success rate of acquiring the target fragment is high. It is a good method for rapid amplification of adjacent flanking sequences of the known sequence.