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Tag1 is an autonomous transposable element that shows somatic excision in both Arabidopsis and tobacco.

机译:Tag1是一种自主转座元件,在拟南芥和烟草中均显示出体细胞切除。

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摘要

Tag1 is a transposable element first identified as an insertion in the CHL1 gene of Arabidopsis. The chl1::Tag1 mutant originated from a plant (ecotype Landsberg erecta) that had been transformed with the maize transposon Activator (Ac), which is distantly related to Tag1. Genomic analysis of untransformed Landsberg erecta plants demonstrated that two identical Tag1 elements are present in the Landsberg erecta genome. To determine what provides transposase function for Tag1 transposition, we examined Tag1 excision in different genetic backgrounds. First, the chl1::Tag1 mutant was backcrossed to untransformed wild-type Arabidopsis plants to remove the Ac element(s) from the genome. F2 progeny that had no Ac elements but still retained Tag1 in the CHL1 gene were identified. Tag1 still excised in these Ac-minus progeny producing CHL1 revertants; therefore, Ac is not required for Tag1 excision. Next, Tag1 was inserted between a cauliflower mosaic virus 35S promoter and a beta-glucuronidase (GUS) marker gene and transformed into tobacco. Transformants showed blue-staining sectors indicative of Tag1 excision. Transgenic tobacco containing a defective Tag1 element, which was constructed in vitro by deleting an internal 1.4-kb EcoRI fragment, did not show blue-staining sectors. We conclude that Tag1 is an autonomous element capable of independent excision. The 35S-GUS::Tag1 construct was then introduced into Arabidopsis. Blue-staining sectors were found in cotyledons, leaves, and roots, showing that Tag1 undergoes somatic excision during vegetative development in its native host.
机译:Tag1是一种转座因子,首先被鉴定为拟南芥CHL1基因的插入。 chl1 :: Tag1突变体起源于一种植物(生态型Landsberg erecta),该植物已被与Tag1远缘相关的玉米转座子激活剂(Ac)转化。未转化的Landsberg erecta植物的基因组分析表明,Landsberg erecta基因组中存在两个相同的Tag1元件。为了确定什么为Tag1转座提供转座酶功能,我们检查了不同遗传背景下的Tag1切除。首先,将chl1 :: Tag1突变体回交到未转化的野生型拟南芥植物中,以从基因组中去除Ac元素。鉴定出没有Ac元件但仍保留Tag1在CHL1基因中的F2后代。 Tag1仍在这些产生Ac负子代的CHL1回复子中切除;因此,Tag1切除不需要Ac。接下来,将Tag1插入花椰菜花叶病毒35S启动子和β-葡糖醛酸糖苷酶(GUS)标记基因之间,然后转化为烟草。转化子显示蓝色染色,指示Tag1切除。含有有缺陷的Tag1元件的转基因烟草,通过删除内部的1.4kb EcoRI片段在体外构建而成,没有显示蓝色。我们得出结论,Tag1是能够独立切除的自主元件。然后将35S-GUS :: Tag1构建体引入拟南芥中。在子叶,叶和根中发现了蓝色染色部分,这表明Tag1在其原生宿主的营养发育过程中经历了体细胞切除。

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