title: Development and application of a general plasmid reference material for GMO screening.
abstract: The use of analytical controls is essential when performing GMO detection through screening tests. Additionally, the presence of taxon-specific sequences is analyzed mostly for quality control during GMO detection. In this study, 11 commonly used genetic elements involving three promoters (P-35S, P-FMV35S and P-NOS), four marker genes (Bar, NPTII, HPT and Pmi), and four terminators (T-NOS, T-35S, T-g7 and T-e9), together with the reference gene fragments from six major crops of maize, soybean, rapeseed, rice, cotton and wheat, were co-integrated into the same single plasmid to construct a general reference plasmid pBI121-Screening. The suitability test of pBI121-Screening plasmid as reference material indicated that the non-target sequence on the pBI121-Screening plasmid did not affect the PCR amplification efficiencies of screening methods and taxon-specific methods. The sensitivity of screening and taxon-specific assays ranged from 5 to 10 copies of pBI121-Screening plasmid, meeting the sensitivity requirement of GMO detection. The construction of pBI121-Screening solves the lack of a general positive control for screening tests, thereby reducing the workload and cost of preparing a plurality of the positive control.
authors: 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: Yuhua
authors: Y
authors: Wu
authors: 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: Jun
authors: J
authors: Li
authors: 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: Yulei
authors: Y
authors: Wang
authors: 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: Xiaofei
authors: X
authors: Li
authors: 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: Yunjing
authors: Y
authors: Li
authors: 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: Li
authors: L
authors: Zhu
authors: 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: Jun
authors: J
authors: Li
authors: 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. Electronic address: wugang@caas.cn.
authors: Gang
authors: G
authors: Wu
mesh_terms: D058893:DNA Barcoding, Taxonomic; D017343:Genes, Plant; D030821:Plants, Genetically Modified / Q000145:classification / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012015:Reference Standards; D012680:Sensitivity and Specificity
keywords: GMO; General positive control; Genetic element; Reference gene; Screening