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Successful detection of foreign inserts in transgenic rice TT51‐1 (BT63) by RNA‐sequencing combined with PCR
Author(s) -
Li Yunjing,
Li Jun,
Wu Yuhua,
Cao Yinglong,
Li Jun,
Zhu Li,
Li Xiaofei,
Huang Shunmou,
Wu Gang
Publication year - 2016
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.7913
Subject(s) - biology , terminator (solar) , gene , dna sequencing , genetics , primer (cosmetics) , genome , computational biology , polymerase chain reaction , oryza sativa , ionosphere , chemistry , physics , organic chemistry , astronomy
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. CONCLUSION 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