title: Successful detection of foreign inserts in transgenic rice TT51-1 (BT63) by RNA-sequencing combined with PCR.
abstract: BACKGROUND
  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.
  
  
  CONCLUSIONS
  The novel approach has a great potential for application to the detection and identification of unauthorised GMOs in food and feed products. © 2016 Society of Chemical Industry.
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Yunjing
authors.initials: Y
authors.lastname: Li
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Yinglong
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Li
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.
authors.forename: Shunmou
authors.initials: S
authors.lastname: Huang
authors.affiliation: Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China.|Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan, 430062, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wu
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D000594:Amino Acid Oxidoreductases / Q000235:genetics; D001413:Bacillus thuringiensis / Q000235:genetics; D030561:Databases, Nucleic Acid; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D013728:Terminator Regions, Genetic
chemical_list: D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase
keywords: PCR; Solexa sequencing; TT51-1; genetically modified organism