您好，欢迎访问浙江省农业科学院 机构知识库！
浙江省农业科学院机构知识库

全部

  • 全部
  • 标题
  • 学者
  • 机构
  • 关键词

TTLOC: A Tn5 transposase-based approach to localize T-DNA integration sites

文献类型： 外文期刊

作者： Tao, Xiao-Yuan 1 ; Feng, Shou-Li 1 ; Li, Xin-Jia 1 ; Li, Yan-Jun 1 ; Wang, Wei 2 ; Gilliham, Matthew 3 ; Chen, Zhong-Hua 4 ; Xu, Sheng-Chun 1 ;

作者机构： 1.Xianghu Lab, Biotechnol Inst, Hangzhou 311231, Peoples R China

2.Nanjing Agr Univ, Collaborat Innovat Ctr Modern Crop Prod CIC MCP, Natl Key Lab Crop Genet & Germplasm Enhancement &, Coll Agr,Zhongshan Biol Breeding Lab, Nanjing 210095, Jiangsu, Peoples R China

3.Univ Adelaide, ARC Ctr Excellence Plants Space, Sch Agr Food & Wine, Waite Research Precinct, Glen Osmond, SA 5064, Australia

4.Western Sydney Univ, Sch Sci, Penrith, NSW 2751, Australia

5.Zhejiang Acad Agr Sci, Inst Digital Agr, Hangzhou 310021, Peoples R China

期刊名称：PLANT PHYSIOLOGY （ 影响因子：6.9； 五年影响因子：7.7 ）

ISSN： 0032-0889

年卷期： 2025 年 197 卷 4 期

页码：

收录情况： SCI

摘要： Thermal asymmetric interlaced-polymerase chain reaction-based and whole-genome sequencing-based T-DNA localization approaches have been developed for the recovery of T-DNA integration sites (TISs). Nevertheless, a low-cost and high-throughput technique for the detection of TISs, which would facilitate the identification of genetically engineered plants, is in high demand for rapid crop breeding and plant synthetic biology. Here, we present Tn5 transposase-based T-DNA integration site localization (TTLOC), a Tn5-based approach for TIS localization. TTLOC employs specialized adaptor-assembled Tn5 transposases for genomic DNA tagmentation. TTLOC library construction is straightforward, involving only six steps that requires two and a half hours to complete. The resulting pooled library is compatible with next-generation sequencing, which enables high-throughput determination. We demonstrate the ability of TTLOC to recover 95 non-redundant TISs from 65 transgenic Arabidopsis (Arabidopsis thaliana) lines, and 37 non-redundant TISs from the genomes of transgenic rice (Oryza sativa), soybean (Glycine max), tomato (Solanum lycopersicum), potato (Solanum tuberosum), and from the large hexaploid wheat (Triticum aestivum) genome. TTLOC is a cost-effective method, as 1 to 2 Gb of raw data for each multiplexing library are sufficient for efficient TIS calling, independent of the genome size. Our results establish TTLOC as a promising strategy for evaluation of genome engineered plants and for selecting genome safe harbors for trait stacking in crop breeding and plant synthetic biology. TTLOC is a Tn5 transposase-based tool for evaluating genetically engineered plants, with potential applications in crop breeding and plant synthetic biology.

  • 相关文献
作者其他论文 更多>>
置顶
购物车
反馈

© 京ICP备09089781号-29 主办单位：浙江省农业科学院

学术资源： 中国农业科学院 农业专业知识服务系统 农科联盟资源共建共享平台 中国农业科技文献与信息服务平台 中国农业期刊集成服务平台