A Survey of New Temperature-Sensitive, Embryonic-Lethal Mutations in C. elegans: 24 Alleles of Thirteen Genes
A Survey of New Temperature-Sensitive, Embryonic-Lethal Mutations in C. elegans: 24 Alleles of Thirteen Genes
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DOI：
10.1371/journal.pone.0016644
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发表时间：
2011-03-01
期刊：
影响因子：
3.7
通讯作者：
Bowerman, Bruce
中科院分区：
文献类型：
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作者：
O'Rourke, Sean M.;Carter, Clayton;Bowerman, Bruce
研究C.在elegans胚胎中，我们筛选了温度敏感的胚胎致死突变，试图绕过幼虫和成虫生殖系发育过程中对这些基因的基本合子要求。对于条件等位基因，可以通过在不同时间从15摄氏度的允许温度转换到26摄氏度的限制温度来检查多种基本要求。在这里，我们描述了24个条件突变，影响13个不同的基因座，并报告的身份的基因突变负责的条件致死突变体中的22个。除了四个突变外，所有突变都是错义突变，其中两个突变影响剪接位点，另一个产生框内缺失，另一个产生过早终止密码子。几乎所有的错义突变都影响直系同源物中保守的残基，因此可能对其他生物体中的工程条件突变有用。我们发现，62%的突变体显示额外的表型时，转移到限制性温度为L1幼虫，除了导致胚胎致死后L4 upshifts。值得注意的是，我们还发现，24个突变中有13个似乎是快速作用的，这使得它们对于仔细分析多种基本需求特别有用。我们的研究结果突出了C的价值。elegans用于识别必需基因中有用的温度敏感性突变，并为一些受影响的基因座的要求提供新的见解。
To study essential maternal gene requirements in the early C. elegans embryo, we have screened for temperature-sensitive, embryonic lethal mutations in an effort to bypass essential zygotic requirements for such genes during larval and adult germline development. With conditional alleles, multiple essential requirements can be examined by shifting at different times from the permissive temperature of 15 degrees C to the restrictive temperature of 26 degrees C. Here we describe 24 conditional mutations that affect 13 different loci and report the identity of the gene mutations responsible for the conditional lethality in 22 of the mutants. All but four are mis-sense mutations, with two mutations affecting splice sites, another creating an in-frame deletion, and one creating a premature stop codon. Almost all of the mis-sense mutations affect residues conserved in orthologs, and thus may be useful for engineering conditional mutations in other organisms. We find that 62% of the mutants display additional phenotypes when shifted to the restrictive temperature as L1 larvae, in addition to causing embryonic lethality after L4 upshifts. Remarkably, we also found that 13 out of the 24 mutations appear to be fast-acting, making them particularly useful for careful dissection of multiple essential requirements. Our findings highlight the value of C. elegans for identifying useful temperature-sensitive mutations in essential genes, and provide new insights into the requirements for some of the affected loci.