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首页> 外文期刊>Acta Physiologiae Plantarum >Cre-mediated marker gene removal for production of biosafe commercial oilseed rape
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Cre-mediated marker gene removal for production of biosafe commercial oilseed rape

机译：Cre-介导的标志物基因去除生物皂卓越商业油菜的生产

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The Cre/loxP-based self-excision represents a promising strategy for removal of hazardous transgene(s) from genomes of targeted crops prior to their introduction into environment. Here, we applied the Cre/loxP self-excision strategy in which the cre recombinase gene is driven by the embryo-specific CRUC promoter from Arabidopsis thaliana. Besides, the Cre/loxP cassette, the T-DNA also consisted of the -glucuronidase gene, controlled by the double dCaMV 35S promoter and the selectable neomycin phosphotransferase gene; the latter was aimed to be removed from the transgenic genome. The Cre/loxP self-excision cassette was introduced into genomes of four commercial oilseed rape cultivars Haydn, Heros, Hunter and Topas (Brassica napus L.) via Agrobacterium tumefaciens. Transgenic T-0 plants were regenerated from all cultivars. Their detailed molecular analyses revealed premature activation of the CRUC promoter that resulted in partial excision of the nptII gene. Progenies of four self-pollinated T-0 lines were further analysed, and the data confirmed complete excision event in 5 out of 105 (4.8%) of T-1 transgenic oilseed rape plants. The excision efficiency does not seem to depend on the target cultivar. However, the poor transformation efficiency of rapeseed and the limited specificity of the CRUC promoter are clearly the bottleneck of this approach, and the feasibility of (tissue-specific) self-excision of selectable marker gene from genomes of each commercial rapeseed variety adds to their perspective to cope the increasing negative impacts of climate changes.
机译：基于CRE / LOXP的自我切除是在进入环境之前从靶向作物的基因组中去除危险转基因的有希望的策略。在这里，我们应用CRE / LOXP自我切除策略，其中CRE重组酶基因由拟南芥拟南芥的胚胎特异性Crauc启动子驱动。此外，CRE / LOXP盒，T-DNA也由葡糖醛酶基因组成，由双DCAMV 35s启动子和可选的新霉素磷酸转移酶基因控制;后者旨在从转基因基因组中除去。通过农杆菌突发，CRE / LOXP自我切除暗盒引入了四种商用油菜品种Haydn，Heros，Hunter和Topas（Brassica Napus L.）的基因组。转基因T-0植物从所有品种中再生。它们的详细分子分析显示了CRUC启动子的过早激活，导致NPTII基因的部分切除。进一步分析了四种自授粉T-0系的后代，数据确认了105个（4.8％）T-1转基因油菜植物中的5个完全切除事件。切除效率似乎不依赖于目标品种。然而，油菜籽的转化效率差和CRUC启动子的有限特异性显然是这种方法的瓶颈，并且来自每个商业油菜籽品种的选择性标记基因的可选标志物基因的可行性增加了它们透视应对气候变化的日益较大的负面影响。

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