数字农科院2.0

Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events

文献类型： 外文期刊

作者： Wu Yu-hua;Zhang Li;Wu Gang;Nie Shu-jing;Lu Chang-ming;Zhang Li

作者机构：

关键词： transgenic rapeseed;junction fragment;pre-insertion site;DNA rearrangement

期刊名称： JOURNAL OF INTEGRATIVE AGRICULTURE

ISSN： 2095-3119

年卷期： 2014 年 13 卷 9 期

页码：

收录情况： SCI

摘要： 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 MS 1, 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.

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