Gomez-Diaz Carolina, Alcorta Esther
Department of Functional Biology, University of Oviedo, Oviedo, Spain.
Genesis. 2008 Jun;46(6):283-8. doi: 10.1002/dvg.20394.
Mosaics have been used in Drosophila to study development and to generate mutant structures when a mutant allele is homozygous lethal. New approaches of directed somatic recombination based on FRT/FLP methods, have increased mosaicism rates but likewise multiple clones in the same individual appeared more frequently. Production of single clones could be essential for developmental studies; however, for cell-autonomous gene function studies only the presence of homozygous cells for the target recessive allele is relevant. Herein, we report the number and extension of antennal mosaics generated by the MARCM system at different ages. This information is directed to obtain the appropriated mosaic type for the intended application. By applying heat shock at 10 different developmental stages from 0-12 h to 6-7 days after egg laying, more than 50% of mosaics were obtained from 5,028 adults. Single recombinant clones appeared mainly at early stages while massive recombinant areas were observed with late treatments.
镶嵌体已被用于果蝇研究发育过程,以及在突变等位基因纯合致死时产生突变结构。基于FRT/FLP方法的定向体细胞重组新方法提高了镶嵌率,但同一个体中出现多个克隆的情况也更频繁。单克隆的产生对于发育研究可能至关重要;然而,对于细胞自主基因功能研究而言,只有目标隐性等位基因的纯合细胞的存在才是相关的。在此,我们报告了由MARCM系统在不同年龄产生的触角镶嵌体的数量和范围。这些信息旨在为预期应用获得合适的镶嵌体类型。通过在产卵后0至12小时到6至7天的10个不同发育阶段施加热休克,从超过5028只成虫中获得了超过50%的镶嵌体。单重组克隆主要出现在早期阶段,而后期处理则观察到大量重组区域。