果蝇卵巢中干细胞谱系的遗传镶嵌分析。
Genetic Mosaic Analysis of Stem Cell Lineages in the Drosophila Ovary.
作者信息
Laws Kaitlin M, Drummond-Barbosa Daniela
机构信息
Division of Reproductive Biology, Department of Biochemistry and Molecular Biology, Johns Hopkins Bloomberg School of Public Health, 615 N. Wolfe Street, Baltimore, MD, 21205, USA.
出版信息
Methods Mol Biol. 2015;1328:57-72. doi: 10.1007/978-1-4939-2851-4_4.
Abstract
Genetic mosaic analyses represent an invaluable approach for the study of stem cell lineages in the Drosophila ovary. The generation of readily identifiable, homozygous mutant cells in the context of wild-type ovarian tissues within intact organisms allows the pinpointing of cellular requirements for gene function, which is particularly important for understanding the physiological control of stem cells and their progeny. Here, we provide a step-by-step guide to the generation and analysis of genetically mosaic ovaries using flippase (FLP)/FLP recognition target (FRT)-mediated recombination in adult Drosophila melanogaster, with a focus on the processes of oogenesis that are controlled by diet-dependent factors.
摘要
基因镶嵌分析是研究果蝇卵巢干细胞谱系的一种极有价值的方法。在完整生物体的野生型卵巢组织背景下产生易于识别的纯合突变细胞，能够精准确定基因功能的细胞需求，这对于理解干细胞及其后代的生理控制尤为重要。在这里，我们提供了一份详细的分步指南，介绍如何在成年黑腹果蝇中利用翻转酶（FLP）/FLP识别靶点（FRT）介导的重组来产生和分析基因镶嵌卵巢，重点关注受饮食依赖性因素控制的卵子发生过程。
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