Identification of transgene insertions in two genetically modified soybeans using high throughput next generation sequencing

作     者：Shouwei Zhang Shi Sun Jing Sun Xiujie Liu Wensheng Hou Tingting Wu Cunxiang Wu Tianfu Han Bingjun Jiang 

作者机构：MOA Key Laboratory of Soybean Biology (Beijing)Institute of Crop ScienceThe Chinese Academy of Agricultural Sciences 

出 版 物：《Oil Crop Science》 (中国油料作物学报(英文版）)

年 卷 期：2018年第3卷第2期

页      面：92-98页

学科分类：09[农学] 

基　　金：supported by the Genetically Modified Organisms Breeding Major Projects of China (2016ZX08011-003) China Agriculture Research System (CARS-04) CAAS Agricultural Science and Technology Innovation Project 

主　　题：genetically modified soybean high methionine transgene insertion identification,next generation sequencing 

摘      要：Genetically modified(GM) organisms are widely adopted. However, their safety assessments and control are still of special concern to the public. Identifying and localizing transgene insertion is an essentially prerequisite step. In this study, 2 independent transgene soybean lines were selected(LB4-AtDCGS-1-20-5-2 and CGS-ZG11) as typical cases. Both lines contained expression cassette of At-DCGS that encoding a feedback-insensitive cystathionine gamma-synthase to produce higher level methionine(Met). LB4-AtDCGS-1-20-5-2 was whole genome sequenced with one paired-end 500 bp library and two mate-paired 1 kb and 2 kb libraries using Illumina HiSeq sequencing platform. CGS-ZG11 was sequenced with only one paired-end 500 bp library. Both genomes were assembled,and 2 scaffold sequences(1 for each line) were screened out by aligning with *** the transgene insertion and its flanking regions in soybean genome were further identified and confirmed by PCR cloning and Sanger sequencing. Results showed that these 2 transgene lines had single copy of inserted transgene. Their transgene insertion contents were identified, which facilitates further safety assessment. These results indicated that genome assembly using high throughput sequencing is a powerful tool for identifying transgene insertions, even with limited knowledge.