在斑马鱼中进行成功的雌核发育单倍体筛选。

Undertaking a successful gynogenetic haploid screen in zebrafish.

作者信息

Layton Judith E

机构信息

The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC, 3050, Australia.

出版信息

Methods Mol Biol. 2009;546:31-44. doi: 10.1007/978-1-60327-977-2_3.

DOI:10.1007/978-1-60327-977-2_3

PMID:19378096

Abstract

Chemical mutagenesis using N-ethyl-N-nitrosourea is the current method of choice for dense mutagenesis in zebrafish. Methods are available for both pre-meiotic and post-meiotic sperm mutagenesis; in this chapter, pre-meiotic mutagenesis is described. Mutated males are crossed with untreated females to create an F1 generation that is heterozygous for the mutations. The F1 females can be screened directly by making haploid embryos using in vitro fertilization (IVF) with ultraviolet (UV)-irradiated sperm. This approach requires substantially fewer fish and less aquarium space than the classical F2 generation screen and is feasible for a small research group. Production of haploid embryos is described in detail.

摘要

使用N-乙基-N-亚硝基脲进行化学诱变是目前在斑马鱼中进行密集诱变的首选方法。有适用于减数分裂前和减数分裂后精子诱变的方法；在本章中，将描述减数分裂前诱变。使突变的雄性与未处理的雌性杂交，以产生对突变杂合的F1代。可以通过使用紫外线（UV）照射的精子进行体外受精（IVF）来制作单倍体胚胎，从而直接筛选F1雌性。与传统的F2代筛选相比，这种方法所需的鱼类和水族箱空间要少得多，并且对于小型研究小组来说是可行的。本文将详细描述单倍体胚胎的产生。