基于高通量测序技术的转基因玉米GM11061分子特征研究  

Molecular characterization of transgenic maize GM11061 based on high-throughput sequencing technology

作　　者：杨迎霞[1] 张冠 王梦梦 陆国清 王倩[1] 陈锐[1] YANG Ying-Xia;ZHANG Guan;WANG Meng-Meng;LU Guo-Qing;WANG Qian;CHEN Rui(Institute of Germplasm Resources and Biotechnology,Tianjin Academy of Agricultural Sciences,Tianjin 300381,China;Tianjin Key Laboratory of Food Biotechnology/School of Biotechnology and Food Science,Tianjin University of Commerce,Tianjin 300134,China)

机构地区：[1]天津市农业科学院种质资源与生物技术研究所,天津300381 [2]天津商业大学生物技术与食品科学学院/天津市食品生物技术重点实验室,天津300134

出　　处：《作物学报》2022年第7期1843-1850,I0001-I0011,共19页Acta Agronomica Sinica

基　　金：农业农村部农业转基因生物安全评价(食用)重点实验室开放课题(SAGM-Y-2020009)资助。

摘　　要：鉴定外源DNA片段在受体基因组中的插入位点、整合序列与拷贝数信息是转基因植物安全评价体系的关键环节。传统的转基因植物分子特征鉴定技术繁杂费力、耗时低效且具有一定的局限性。本研究利用基因组重测序技术与实验室自建的数据分析流程,针对转基因玉米GM11061开展了分子特征鉴定研究,结果表明:外源DNA片段仅插入受体基因组一个拷贝,位于5号染色体198,621,57~198,621,620 bp之间,不含载体骨架序列,并通过Sanger测序验证了其上下游结合位点。测序数据量梯度分析显示,最低~5×的重测序原始数据可实现该整合位点的鉴定。本研究证实Illumina高通量测序技术与配套的数据分析方法结合可实现简易、快速、精准的植物分子特征鉴定研究。本研究结果有助于植物功能基因组学基础研究,同时也为转基因安全评价体系的完善提供技术支撑。Insertion site,integrated sequences,and copy number information of exogenous DNA integrations in host genome are key steps in the safety evaluation of genetically modified plants.Traditional identification techniques for molecular characteristics of transgenic plants are complicated,laborious,inefficient and limited.Here,we reported the molecular characterization of one new transgenic maize event GM11061 via a whole genome(paired-end)sequencing approach and laboratory-developed bioinfor-matics methods.The results showed there was only one copy of the exogenous DNA inserted,which was located within the region of 198,621,571–198,621,620 bp on chromosome 5 in GM11061 genome,without the vector skeleton sequence.Its upstream and downstream binding sites were validated using conventional PCR and sanger sequencing.Gradient analysis of sequencing data revealed that the lowest 5×resequencing raw data could be used to identify the integration site.The study confirmed that the whole genome resequencing approach combined with bioinformatics methods could achieve simple,rapid,and accurate identifi-cation of plant molecular characteristics.These results are helpful to the basic research of functional genomics and provide technical support for the safety evaluation of transgenic plants.

关 键 词：高通量测序 转基因玉米 分子特征 

分 类 号：S513[农业科学—作物学]