Muñoz Manuel J, Riddle Donald L
Molecular Biology Program and Division of Biological Sciences, University of Missouri, Columbia, Missouri 65211-7400, USA.
Genetics. 2003 Jan;163(1):171-80. doi: 10.1093/genetics/163.1.171.
We developed selective conditions for long-lived mutants of the nematode Caenorhabditis elegans by subjecting the first larval stage (L1) to thermal stress at 30 degrees for 7 days. The surviving larvae developed to fertile adults after the temperature was shifted to 15 degrees. A total of one million F(2) progeny and a half million F(3) progeny of ethyl-methanesulfonate-mutagenized animals were treated in three separate experiments. Among the 81 putative mutants that recovered and matured to the reproductive adult, 63 retested as thermotolerant and 49 (80%) exhibited a >15% increase in mean life span. All the known classes of dauer formation (Daf) mutant that affect longevity were found, including six new alleles of daf-2, and a unique temperature-sensitive, dauer-constitutive allele of age-1. Alleles of dyf-2 and unc-13 were isolated, and mutants of unc-18, a gene that interacts with unc-13, were also found to be long lived. Thirteen additional mutations define at least four new genes.
我们通过将线虫秀丽隐杆线虫的第一幼虫期（L1）在30摄氏度下热应激处理7天，为其开发了长寿突变体的筛选条件。温度转移到15摄氏度后，存活的幼虫发育成可育成虫。在三个独立实验中，对总共100万个经甲基磺酸乙酯诱变的动物的F(2)后代和50万个F(3)后代进行了处理。在81个恢复并成熟到生殖成虫阶段的假定突变体中，63个经重新测试具有耐热性，49个（80%）平均寿命增加超过15%。发现了所有已知的影响寿命的 dauer 形成（Daf）突变体类别，包括六个新的daf-2等位基因，以及一个独特的温度敏感、 dauer 组成型age-1等位基因。分离出了dyf-2和unc-13的等位基因，并且还发现与unc-13相互作用的基因unc-18的突变体寿命也很长。另外13个突变定义了至少四个新基因。
