Wang Xu-Jing, Jiao Yue, Ma Shuo, Yang Jiang-Tao, Wang Zhi-Xing
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences/MARA Key Laboratory on Safety Assessment (Molecular) of Agri-GMO, Beijing China.
Development Center for Science and Technology/MARA, Beijing, China.
Front Plant Sci. 2020 Dec 1;11:573871. doi: 10.3389/fpls.2020.573871. eCollection 2020.
Molecular characterization is a key step in the risk assessment of genetically modified organisms (GMOs) for regulatory approval. Herein, we describe a method for analyzing copy number, insertion loci, and flanking sequences through whole-genome sequencing (WGS) and bioinformatics. Comprehensive molecular characterization of G2-6 transgenic rice was performed using this pipeline. The results showed that one copy of the foreign gene was inserted into rice chromosome 8. There was no vector backbone insertion but an unexpected insertion and DNA rearrangement at the 3' end of the T-DNA. We also obtained the 5' and 3' flanking sequences of the T-DNA. Our results suggested that the use of a combination of WGS and bioinformatics is an effective strategy for the molecular characterization of GMOs.
分子特征分析是转基因生物(GMO)进行监管审批风险评估的关键步骤。在此,我们描述了一种通过全基因组测序(WGS)和生物信息学分析拷贝数、插入位点及侧翼序列的方法。使用该流程对G2-6转基因水稻进行了全面的分子特征分析。结果表明,外源基因的一个拷贝插入到水稻第8号染色体。没有载体骨架插入,但在T-DNA的3'端存在意外插入和DNA重排。我们还获得了T-DNA的5'和3'侧翼序列。我们的结果表明,结合使用WGS和生物信息学是转基因生物分子特征分析的有效策略。