Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events
To characterize the DNA rearrangement of both the T-DNA region and the genomic insertion site during T-DNA insertion, the Genomewalker strategy was used to isolate the junctions between the inserted DNA and the plant genomic DNA in six rapeseed events as well as the genomic DNA at the sites before i...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
KeAi Communications Co., Ltd.
2014-09-01
|
| Series: | Journal of Integrative Agriculture |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311913606280 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849253382066274304 |
|---|---|
| author | Yu-hua WU Li ZHANG Gang WU Shu-jing NIE Chang-ming LU |
| author_facet | Yu-hua WU Li ZHANG Gang WU Shu-jing NIE Chang-ming LU |
| author_sort | Yu-hua WU |
| collection | DOAJ |
| description | To characterize the DNA rearrangement of both the T-DNA region and the genomic insertion site during T-DNA insertion, the Genomewalker strategy was used to isolate the junctions between the inserted DNA and the plant genomic DNA in six rapeseed events as well as the genomic DNA at the sites before integration. During transformation in each of the six events, portions of both the right border (RB) and left border (LB) regions of the T-DNA were deleted, ranging from a 7 nucleotide deletion of the LB repeats in event RF1 to a 207 bp deletion of the LB region in event RF2. For the six events, T-DNA integration resulted in a deletion at the target site spanning less than 100 bp. Sequence analysis indicated that the T-DNA was integrated into the coding region of various native rapeseed genes in events RF1 and RF2. Duplications of the genomic DNA target site were observed in events RF2, RF3 and Topas 19/2. And multimerization of transgenes was found in event Topas 19/2, in which, the T-DNA was integrated as a head-to-head (RB-to-RB) concatemer into the recipient genome. In event MS1, chromosomal translocation or a large target-site deletion may have occurred during T-DNA integration, which was identified due to a failure to amplify the presumptive insertion site based on the flanking rapeseed DNA sequences. Our results provide comprehensive data concerning transgene organization and the genomic context of the T-DNA in six rapeseed events, which can aid in the developing of insert fingerprinting and the monitoring of long-term genetic stability and potential unintended effects of transgenic events. |
| format | Article |
| id | doaj-art-568ba64098a7426f869592a64498a24f |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2014-09-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-568ba64098a7426f869592a64498a24f2025-08-20T03:56:19ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192014-09-011391865187610.1016/S2095-3119(13)60628-0Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed EventsYu-hua WU0Li ZHANG1Gang WU2Shu-jing NIE3Chang-ming LU4Oilcrops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, P.R. China; WU Yu-hua, Tel: +86-27-86711501; ZHANG LiOilcrops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, P.R. China; College of Life Science, South-Central University for Nationalities, Wuhan 430074, P.R. China; WU Yu-hua, Tel: +86-27-86711501; ZHANG LiOilcrops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, P.R. ChinaOilcrops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, P.R. ChinaOilcrops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, P.R. China; Correspondence LU Chang-ming, Tel: +86-27-86728186, Fax: +86-27-86711573To characterize the DNA rearrangement of both the T-DNA region and the genomic insertion site during T-DNA insertion, the Genomewalker strategy was used to isolate the junctions between the inserted DNA and the plant genomic DNA in six rapeseed events as well as the genomic DNA at the sites before integration. During transformation in each of the six events, portions of both the right border (RB) and left border (LB) regions of the T-DNA were deleted, ranging from a 7 nucleotide deletion of the LB repeats in event RF1 to a 207 bp deletion of the LB region in event RF2. For the six events, T-DNA integration resulted in a deletion at the target site spanning less than 100 bp. Sequence analysis indicated that the T-DNA was integrated into the coding region of various native rapeseed genes in events RF1 and RF2. Duplications of the genomic DNA target site were observed in events RF2, RF3 and Topas 19/2. And multimerization of transgenes was found in event Topas 19/2, in which, the T-DNA was integrated as a head-to-head (RB-to-RB) concatemer into the recipient genome. In event MS1, chromosomal translocation or a large target-site deletion may have occurred during T-DNA integration, which was identified due to a failure to amplify the presumptive insertion site based on the flanking rapeseed DNA sequences. Our results provide comprehensive data concerning transgene organization and the genomic context of the T-DNA in six rapeseed events, which can aid in the developing of insert fingerprinting and the monitoring of long-term genetic stability and potential unintended effects of transgenic events.http://www.sciencedirect.com/science/article/pii/S2095311913606280transgenic rapeseedjunction fragmentpre-insertion siteDNA rearrangement |
| spellingShingle | Yu-hua WU Li ZHANG Gang WU Shu-jing NIE Chang-ming LU Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events Journal of Integrative Agriculture transgenic rapeseed junction fragment pre-insertion site DNA rearrangement |
| title | Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events |
| title_full | Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events |
| title_fullStr | Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events |
| title_full_unstemmed | Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events |
| title_short | Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events |
| title_sort | characterization of genomic integration and transgene organization in six transgenic rapeseed events |
| topic | transgenic rapeseed junction fragment pre-insertion site DNA rearrangement |
| url | http://www.sciencedirect.com/science/article/pii/S2095311913606280 |
| work_keys_str_mv | AT yuhuawu characterizationofgenomicintegrationandtransgeneorganizationinsixtransgenicrapeseedevents AT lizhang characterizationofgenomicintegrationandtransgeneorganizationinsixtransgenicrapeseedevents AT gangwu characterizationofgenomicintegrationandtransgeneorganizationinsixtransgenicrapeseedevents AT shujingnie characterizationofgenomicintegrationandtransgeneorganizationinsixtransgenicrapeseedevents AT changminglu characterizationofgenomicintegrationandtransgeneorganizationinsixtransgenicrapeseedevents |