Evidence for Drosophila P element transposase activity in mammalian cells and yeast.
Evidence for Drosophila P element transposase activity in mammalian cells and yeast.
复制标题
哺乳动物细胞和酵母中果蝇 P 元件转座酶活性的证据。
DOI：
10.1016/0022-2836(88)90250-1
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发表时间：
1988
影响因子：
5.6
通讯作者：
Rubin,GM
中科院分区：
文献类型：
--
作者：
Rio,DC;Barnes,G;Laski,FA;Rine,J;Rubin,GM
果蝇P元件转座酶的表达是有限的生殖细胞的组织特异性剪接的三个内含子之一。通过体外诱变去除该内含子，可以在果蝇体细胞组织中检测到P元件切除和转座。为了确定P元件转座酶是否可以在其他生物中发挥作用，我们分别在哺乳动物细胞和酵母中表达了缺少一个内含子或缺少所有三个内含子的修饰的P元件。使用P元件切除的测定，我们在培养的猴细胞中检测到明显的切除事件。此外，完整的P元件cDNA的表达对携带RAD 52基因突变的酿酒酵母细胞是致命的，这表明可能通过转座酶作用产生双链DNA断裂。
DrosophilaP element transposase expression is limited to the germline by tissue-specific splicing of one of its three introns. Removal of this intron by mutagenesisin vitrohas allowed both P element excision and transposition to be detected inDrosophilasomatic tissues. In order to determine if P element transposase can function in other organisms, we have expressed modified P elements either lacking one intron or lacking all three introns in mammalian cells and yeast, respectively. Using an assay for P element excision, we have detected apparent excision events in cultured monkey cells. Furthermore, expression of the complete P element cDNA is lethal toSaccharomyces cerevisiaecells carrying a mutation in theRAD52 gene, indicating that double-stranded DNA breaks are generated, presumably by transposase action.