摘要
[目的]建立MON89788大豆转化体特异性定性PCR检测方法。[方法]利用TAIL-PCR技术分离MON89788大豆的3′端旁侧序列,据此序列设计特异性引物进行PCR检测,并对该方法的特异性和灵敏度进行测试。[结果]获得了1142bp的3′端旁侧序列;依据该序列建立的PCR检测方法能特异性从MON89788大豆中扩增出170bp的产物,检测灵敏度达到0.05%,约为40个起始模板拷贝。[结论]该研究建立的方法特异性强、灵敏度高,可适用于MON89788大豆检测。
[Objective] The aim was to establish an event-specific qualitative PCR method for transgenic soybean MON89788. [Method] Firstly,the 3’-junction sequence between host plant DNA and integrated DNA of transgenic MON89788 soybean was isolated using thermal asymmetric interlaced-PCR （TAIL-PCR）,and the specific PCR primers were designed based on the 3’-junction sequence. Secondly,the specificity and sensitivity of the qualitative PCR detection methods employing these primers were tested. [Result] 1142-bp 3’-junction sequence was obtained. According to the sequence,event-specific qualitative PCR method was established,amplifying a 170-bp product specifically from MON89788 event,and the limit of detection was 0.05%,approximately 40 initial template copies. [Conclusion] The method was highly specific,sensitive,and suitable for detection of MON89788 event.
出处
《安徽农业科学》
CAS
北大核心
2010年第13期6679-6682,共4页
Journal of Anhui Agricultural Sciences
基金
国家转基因生物新品种培育重大专项(2008ZX08012-001)
