Small-scale marker-based screening for mutations in zebrafish development.
Small-scale marker-based screening for mutations in zebrafish development.
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基于标记的小规模斑马鱼发育突变筛查。
DOI：
10.1007/978-1-60327-483-8_34
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Ingham,PhilipW
中科院分区：
文献类型：
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作者：
Currie,PeterD;Schilling,ThomasF;Ingham,PhilipW
我们描述了一个标准化的诱变方案和方法，小规模的定向筛选斑马鱼基因组的突变在特定的发展过程。这些方法主要基于在Tubingen，德国、Boston，MA和尤金，OR开发的大型筛选方法以及我们在较小设施中的经验。通过将基于标记的筛选方案与单倍体和二倍体筛选方法相结合，可以在特定过程中有效地回收突变体。随机诱变提供了无偏倚地调查生物体基因组的能力，以寻找在特定过程中起作用的基因。多年来，遗传学家一直在收获通过这种诱变方法产生的突变的丰富收获，针对果蝇Drosophila melanogaster的特定发育过程（1）。对这种方法所揭示的基因的分析彻底改变了我们对动物发育遗传控制的理解，研究人员渴望看到类似的诱变方法应用于脊椎动物基因组，但由于经典脊椎动物发育模型的遗传难治性而受阻。小鼠是唯一一种进行了大规模突变筛查的脊椎动物。这些筛选主要限于鉴定出生后可见形态学性状的缺陷，因为胚胎突变表型的筛选很困难，因为发育发生在子宫内（2）。对使用诱变方法研究脊椎动物发育早期方面感兴趣的生物学家被迫寻找替代方法。
We describe a standardized mutagenic protocol and a methodology for small-scale directed screening of the zebrafish genome for mutations in specific developmental processes. The methods are based primarily on those developed for large-scale screens in Tubingen, Germany; Boston, MA; and Eugene, OR as well as our experiences with a smaller facility. By combining a marker-based screening protocol with both haploid and diploid screening methods, one can efficiently recover mutants in specific processes. Random mutagenesis provides the ability to survey the genome of an organism, without bias, for genes that function in particular processes. For many years, geneticists have been reaping the rich harvest of mutations produced by such a mutagenic approach, directed against particular developmental processes of the fruit fly Drosophila melanogaster (1). Analysis of the genes uncovered by this approach has revolutionized our understanding of the genetic control of animal development.Researchers eager to see a similar mutagenic approach applied to the vertebrate genome have been stymied by the genetic intractability of classical vertebrate developmental models. The mouse is the only vertebrate organism in which large-scale screens for mutations have been performed. These screens have, in the main, been limited to identification of defects in visible morphological traits after birth, since screening for embryonic mutant phenotypes is difficult because development occurs in utero (2). Biologists interested in using a mutagenic approach to study early aspects of vertebrate development have been forced to search for an alternative.