中文摘要：由于大多数未经授权转基因生物的事件特异性序列信息目前仍不可用，因此检测未经授权的转基因生物仍然具有挑战性。本文以抗虫水稻TT51-1为例，开发了检测转基因植株的RNA测序与PCR技术的结合方法。研究发现，使用数据库的相似性分析可以识别测试样本。结果表明，基于RNA测序与PCR技术的检测方法具有广阔的应用前景，可用于食品和饲料中未经授权的转基因食品的检测和鉴定。
外文摘要： As event-specific sequence information for most unauthorised genetically modified organisms (GMOs) is currently still unavailable, detecting unauthorised GMOs remains challenging. Here, we used insect-resistant rice TT51-1 as an example to develop a novel approach via detecting GMOs by RNA-seq (sequencing) and PCR. RNA-seq of TT51-1 generated 4.8 million (M) 21-nt cDNA tags. Alignment to the Oryza sativa subsp. japonica reference genome revealed 24 098 unmapped tags. Foreign tags from the nopaline synthetic enzyme gene (NOS) terminator and insect-resistant genes were then identified by searching against the NCBI VecScreen and NT databases. RESULTS: To further isolate foreign DNA sequences, putative NOS terminator and insect-resistant gene tags were combined and used directly as primer pairs for long-range PCR, producing a 5016-bp fragment. The inserted DNA sequence of TT51-1 has been submitted to a database, and thus, similarity analysis using the database could identify a test sample. CONCLUSION: The novel approach has a great potential for application to the detection and identification of unauthorised GMOs in food and feed products. 
外文关键词： genetically modified organism, TT51-1, Solexa sequencing, PCR
作者：Li, Yunjing; Li, Jun; Wu, Yuhua; Cao, Yinglong; Zhu, Li; Li, Xiaofei; Huang, Shunmou; Wu, Gang
作者单位：Chinese Acad Agr Sci
期刊名称：JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
期刊影响因子：1.815
出版年份：2018
出版刊次：5
点击下载：基于RNA测序与PCR技术的转基因水稻TT51-1（BT63）外源插入基因的成功检测研究