SINGLE-STEP SELECTION FOR TY1 ELEMENT RETROTRANSPOSITION
SINGLE-STEP SELECTION FOR TY1 ELEMENT RETROTRANSPOSITION
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DOI：
10.1073/pnas.88.3.936
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发表时间：
1991-02-01
影响因子：
11.1
通讯作者：
GARFINKEL, DJ
中科院分区：
文献类型：
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作者：
CURCIO, MJ;GARFINKEL, DJ
酵母反转录转座子Ty1被标记了一个报告基因，该基因可以选择RNA介导的转座事件，并适用于其他生物中反转录元件的研究。报告基因是酵母HIS3基因，在反义方向上被人工内含子(AI)打断。HIS3AI序列被插入到Ty1元件中，使得内含子位于Ty1元件的正义链上；因此，标记的Ty1转录本的剪接和逆转录转座可以产生His+细胞。将Ty1-H3mHIS3AI元件与可诱导的GAL1启动子融合，可在半乳糖诱导下产生高频率的组氨酸原营养体。此外，来自含有基因组TymHIS3AI元件的菌株的自发His+逆转株是逆转录转座的结果。通过这种方法，我们估计每代Ty1元件的Ty1转座速率在3×10(-7)到1×10(-5)之间。单个Ty1元素的转座速率的变化与其转录本的相对丰度有关。
The yeast retrotransposon Ty1 has been tagged with a reporter gene that allows selection of RNA-mediated transposition events and is applicable to the study of retroelements in other organisms. The reporter gene is a yeast HIS3 gene interrupted by an artificial intron (AI) in the antisense orientation. The HIS3AI sequences were inserted into a Ty1 element such that the intron is on the sense strand of the Ty1 element; therefore, splicing and retrotransposition of marked Ty1 transcripts can give rise to His+ cells. Fusion of the Ty1-H3mHIS3AI element to the inducible GAL1 promoter resulted in a high frequency of histidine prototrophs upon galactose induction. Moreover, spontaneous His+ revertants derived from strains containing genomic TymHIS3AI elements are a result of retrotransposition. By using this assay, we estimated the Ty1 transposition rate to be between 3 X 10(-7) and 1 X 10(-5) transposition per Ty1 element per generation. Variations in the transposition rate of individual Ty1 elements are correlated with the relative abundance of their transcripts.