Rapid recombination mapping for high-throughput genetic screens in Drosophila.
Rapid recombination mapping for high-throughput genetic screens in Drosophila.
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DOI：
10.1534/g3.113.008615
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发表时间：
2013-12-09
期刊：
影响因子：
--
通讯作者：
Bashirullah A
中科院分区：
文献类型：
--
作者：
Sapiro AL;Ihry RJ;Buhr DL;Konieczko KM;Ives SM;Engstrom AK;Wleklinski NP;Kopish KJ;Bashirullah A
诱变筛选是经典遗传学的主要内容。然而，化学诱导的突变通常难以识别且耗时。在这里，我们报告用显性可见标记对进行重组分析提供了一种快速可靠的策略来绘制果蝇突变图谱。该方法仅需要两代，在小瓶中总共进行六次杂交，就可以高精度地估计负责病变的遗传图谱位置。然后，可以通过平均九个缺陷的互补测试和桑格测序，可靠地使用该遗传图谱位置来识别突变基因。我们已经使用这种方法成功地绘制了第三条染色体上基于甲磺酸乙酯的诱变筛选的一系列突变。我们建议该方法也可以与全基因组测序结合使用，特别是当突变位点的多个独立等位基因不可用时。通过促进突变基因的快速识别，我们的作图策略消除了广泛使用强大的化学诱变筛选来了解基本生物现象的主要障碍。
Mutagenesis screens are a staple of classical genetics. Chemical-induced mutations, however, are often difficult and time-consuming to identify. Here, we report that recombination analysis with pairs of dominant visible markers provides a rapid and reliable strategy to map mutations in Drosophila melanogaster. This method requires only two generations and a total of six crosses in vials to estimate the genetic map position of the responsible lesion with high accuracy. This genetic map position can then be reliably used to identify the mutated gene through complementation testing with an average of nine deficiencies and Sanger sequencing. We have used this approach to successfully map a collection of mutations from an ethyl methanesulfonate−based mutagenesis screen on the third chromosome. We propose that this method also may be used in conjunction with whole-genome sequencing, particularly when multiple independent alleles of the mutated locus are not available. By facilitating the rapid identification of mutated genes, our mapping strategy removes a primary obstacle to the widespread use of powerful chemical mutagenesis screens to understand fundamental biological phenomena.
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影响因子：
12.3
作者：
Cook RK;Christensen SJ;Deal JA;Coburn RA;Deal ME;Gresens JM;Kaufman TC;Cook KR
通讯作者：
Cook KR
影响因子：
3.3
作者：
Ryder, Edward;Ashburner, Michael;Russell, Steven
通讯作者：
Russell, Steven
影响因子：
30.8
作者：
Parks, AL;Cook, KR;Francis-Lang, HL
通讯作者：
Francis-Lang, HL
影响因子：
--
作者：
Sturtevant, AH
通讯作者：
Sturtevant, AH
影响因子：
14.9
作者：
Marygold SJ;Leyland PC;Seal RL;Goodman JL;Thurmond J;Strelets VB;Wilson RJ;FlyBase consortium
通讯作者：
FlyBase consortium