Efficient induction of new mutations at defined loci of the zebrafish

在斑马鱼的特定位点有效诱导新突变

• 批准号: 9420984
• 负责人: David Grunwald
• 金额: $ 38.3万
• 依托单位: University of Utah
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-02-15 至 1999-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9420984&HistoricalAwards=false
• 关键词: Efficient; induction; new; mutations; defined

9420984 Grunwald A new method for mutagenizing zebra fish (Danio rerio) will be developed and optimized. Mutations will be induced in mature sperm with ethylnitrosourea, a potent point mutagen. Founder zebrafish derived from such mutagenized sperm are a rich source of new mutations and can be selected by a failure to complement pre-existing mutations. There are three features of this method which make it a potentially powerful source of new mutations: Germ lines will harbor independent mutations at a frequency of 1-2% at any pre-specified locus; the germ lines of the founders are genetically mosaic; preliminary studies indicate that most of the induced mutations are intragenic mutations. The goals of this proposal are to optimize the mutagenesis procedures and to establish the frequency of germ line specific locus mutations in the founder population; to demonstrate that new and informative mutant alleles of a developmentally important gene can be recovered; and to recover and analyze mutations that affect single genes encompassed by a deletion mutation. %%% Zerbrafish are rapidly becoming important experimental organisms for the study of development. However, while their are many advantages to their use, one serious disadvantage is the lack of an easily manipulated genetic system. The development of a method to generate new and different developmental mutants for analysis will serve the purpose of optimizing this system . ***

9420984 Grunwald拟开发并优化一种新的斑马鱼（Danio rerio）诱变方法。用乙基亚硝基脲（一种强效点诱变剂）在成熟精子中诱导突变。来自这种诱变精子的创始人斑马鱼是新突变的丰富来源，并且可以通过未能补充预先存在的突变来选择。这种方法有三个特点，使其成为一个潜在的强大的新突变来源：生殖系将在任何预先指定的位点以1-2%的频率携带独立的突变;创始人的生殖系是遗传嵌合体;初步研究表明，大多数诱导的突变是基因内突变。 本提案的目标是优化诱变程序，并建立在创始人人口的生殖系特异性基因座突变的频率，以证明，新的和信息突变等位基因的发育重要的基因可以恢复，并恢复和分析突变，影响单个基因所涵盖的缺失突变。 斑马鱼正迅速成为研究发育的重要实验生物。然而，尽管它们的使用有许多优点，但一个严重的缺点是缺乏易于操纵的遗传系统。开发一种产生新的和不同的发育突变体用于分析的方法将有助于优化该系统的目的。 ***