Yang Litao, Wang Congmao, Holst-Jensen Arne, Morisset Dany, Lin Yongjun, Zhang Dabing
1] Collaborative Innovation center for biosafety of GMOs, National Center for Molecular Characterization of GMOs, School of Life Science and Biotechnology, Shanghai Jiao Tong University. 800 Dongchuan Road, Shanghai 200240. P. R. China [2].
Sci Rep. 2013 Oct 3;3:2839. doi: 10.1038/srep02839.
Detection methods and data from molecular characterization of genetically modified (GM) events are needed by stakeholders of public risk assessors and regulators. Generally, the molecular characteristics of GM events are incomprehensively revealed by current approaches and biased towards detecting transformation vector derived sequences. GM events are classified based on available knowledge of the sequences of vectors and inserts (insert knowledge). Herein we present three insert knowledge-adapted approaches for characterization GM events (TT51-1 and T1c-19 rice as examples) based on paired-end re-sequencing with the advantages of comprehensiveness, accuracy, and automation. The comprehensive molecular characteristics of two rice events were revealed with additional unintended insertions comparing with the results from PCR and Southern blotting. Comprehensive transgene characterization of TT51-1 and T1c-19 is shown to be independent of a priori knowledge of the insert and vector sequences employing the developed approaches. This provides an opportunity to identify and characterize also unknown GM events.
公共风险评估者和监管机构的利益相关者需要转基因事件分子特征的检测方法和数据。一般来说,当前方法并不能全面揭示转基因事件的分子特征,且偏向于检测转化载体衍生序列。转基因事件是根据载体和插入片段的序列现有知识(插入片段知识)进行分类的。在此,我们提出了三种基于配对末端重测序的、适用于插入片段知识的转基因事件特征分析方法(以TT51-1和T1c-19水稻为例),具有全面性、准确性和自动化的优点。与PCR和Southern杂交结果相比,发现了两个水稻事件的额外意外插入,从而揭示了其全面的分子特征。采用所开发的方法,TT51-1和T1c-19的全面转基因特征分析显示与插入片段和载体序列的先验知识无关。这为识别和表征未知转基因事件提供了机会。