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诱导降解剂量补偿蛋白 DPY-27 有助于分离出雄性秀丽隐杆线虫进行分子和生化分析。

Inducible degradation of dosage compensation protein DPY-27 facilitates isolation of Caenorhabditis elegans males for molecular and biochemical analyses.

机构信息

Department of Molecular and Cellular Biology, University of California, Davis, Davis, CA 95616, USA.

Biochemistry, Molecular, Cellular and Developmental Biology Graduate Group, University of California, Davis, Davis, CA 95616, USA.

出版信息

G3 (Bethesda). 2022 May 6;12(5). doi: 10.1093/g3journal/jkac085.

Abstract

Biological sex affects numerous aspects of biology, yet how sex influences different biological processes have not been extensively studied at the molecular level. Caenorhabditis elegans, with both hermaphrodites (functionally females as adults) and males, is an excellent system to uncover how sex influences physiology. Here, we describe a method to isolate large quantities of C. elegans males by conditionally degrading DPY-27, a component of the dosage compensation complex essential for hermaphrodite, but not male, development. We show that germ cells from males isolated following DPY-27 degradation undergo meiosis and spermiogenesis like wild type and these males are competent to mate and sire viable offspring. We further demonstrate the efficacy of this system by analyzing gene expression and performing affinity pull-downs from male worm extracts.

摘要

生物性别会影响生物学的众多方面,但性别如何影响不同的生物学过程在分子水平上还没有得到广泛研究。秀丽隐杆线虫既有雌雄同体(成年后具有功能性雌性)又有雄性,是揭示性别如何影响生理机能的绝佳系统。在这里,我们描述了一种通过条件性降解 DPY-27 来分离大量雄性线虫的方法,DPY-27 是剂量补偿复合物的一个组成部分,对于雌雄同体的发育是必需的,但对于雄性的发育不是必需的。我们发现,在 DPY-27 降解后分离出来的雄性生殖细胞能够进行减数分裂和精子发生,与野生型相似,并且这些雄性具有交配和生育有活力后代的能力。我们进一步通过分析基因表达和从雄性虫体提取物中进行亲和下拉实验来证明该系统的有效性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ad1/9073673/0fe1c877951c/jkac085f1.jpg

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