Minos as a genetic and genomic tool in Drosophila melanogaster
Minos as a genetic and genomic tool in Drosophila melanogaster
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DOI：
10.1534/genetics.105.041848
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发表时间：
2005-10-01
期刊：
影响因子：
3.3
通讯作者：
Savakis, C
中科院分区：
文献类型：
--
作者：
Metaxakis, A;Oehler, S;Savakis, C
许多关于黑腹果蝇基因功能的信息都来自于P元素的突变。然而，P元件的主要缺点是它强烈倾向于插入某些基因(热点)，而不插入其他基因(冷点)。在基因中，5‘-UTRs是优先靶点。为了成功地完成果蝇基因组破坏计划，除了P以外的转座子载体的兴起是必要的。我们在这里研究了从果蝇基因组中提取的Minos元件作为果蝇基因组学工具的适宜性。先前的工作表明，Minos是Tc1/Mariner转座元件家族的成员，在不同的生物体和培养细胞中都很活跃；它在几个昆虫物种的胚系、小鼠和人类细胞中产生稳定的整合子。我们产生并分析了96个整合到果蝇基因组中的Minos，并设计了一种有效的产生单一插入的“跳跃启动”方案。插入基因的比例与基因间DNA的比例符合随机分布。在基因内部，有一种统计上显著的偏好，即插入内含子而不是外显子。大约30%的插入是在内含子中，类似于55%的插入是在迄今尚未被P元件击中的基因中或其附近。与其他转座子相比，插入位点表现出对TA二核苷酸插入靶点以外的序列要求很少。我们进一步证明，诱导的Minos插入的再动员可以删除附近的序列。我们的结果表明，Minos是一种补充P元件的有用工具，可用于果蝇的插入突变和基因组分析。
Much of the information about the function of D. melanogaster genes has come from P-element mutagenesis. The major drawback of the P element, however, is its strong bias for insertion into some genes (hotspots) and against insertion into others (coldspots). Within genes, 5'-UTRs are preferential targets. For the Successful completion of the Drosophila Genome Disruption Project, the rise of transposon vectors other than P will be necessary We examined here the suitability of the Minos element from Drosophila hydei as a tool for Drosophila genomics. Previous work has shown that Minos, a member of the Tc1/mariner family of transposable elements, is active in diverse organisms and cultured cells; it produces stable integrants in the germ line of several insect species, in the mouse, and in human cells. We generated and analyzed 96 Minos integrations into the Drosophila genome and devised an efficient 'jump-starting" scheme for production of single insertions. The ratio of insertions into genes vs. intergenic DNA is consistent with a random distribution. Within genes, there is a statistically significant preference for insertion into introns rather than into exons. About 30% of all insertions were in introns and similar to 55% of insertions were into or next to genes that have so far not been hit by the P element. The insertion sites exhibit, in contrast to other transposons, little sequence requirement beyond the TA dinucleotide insertion target. We further demonstrate that induced remobilization of Minos insertions can delete nearby sequences. Our results suggest that Minos is a useful tool complementing the P element for insertional mutagenesis and genomic analysis in Drosophila.