转基因大豆MON89788转化体特异性定性PCR检测（摘要）（英文）

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3条特异性引物用于旁侧序列的扩增。经过3轮PCR扩增,获得特异性扩增产物,将此产物回收后克隆到pMD-18T载体上测序,所得序列用Vector NTI分析,并提交NCBI进行序列比对。并对该方法的特异性和灵敏度进行测试。结果]获得了1 142 bp的3′端旁侧序列。经Blast检索比对,该序列包括2部分,1 ～618 bp为载体序列（E9终止子部分序列和LB序列）,619 ～1 142 bp为大豆基因组序列;依据该序列建立的PCR检测方法能特异性地从MON89788大豆中扩增出170 bp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。

Establishment of Event-specific Qualitative PCR Method for Genetically Modified Soybean MON 89788

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] 1 142-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.