Duhon S A, Murakami S, Johnson T E
Institute for Behavioral Genetics, University of Colorado, Boulder 80309, USA.
Dev Genet. 1996;18(2):144-53. doi: 10.1002/(SICI)1520-6408(1996)18:2<144::AID-DVG7>3.0.CO;2-9.
We have isolated several new EMS-induced, long-lived mutants of Caenorhabditis elegans, using a novel screen that eliminates the need for replica plating. Three new alleles of age-1 (z10, z12, and z25) were identified by failure to complement age-1 (hx546) for life span extension; these alleles had life spans ranging from 18.9 to 25.9 days at 25 degrees C, with an average 46% increase in life span. After backcrossing, alleles were examined in a wild-type background for resistance to several environmental stresses: heat (35 degrees C), ultraviolet (UV) light (20 J/m2), and hydrogen peroxide (H2O2) (0.5 M). Two replicates of the test of thermotolerance were completed on each strain, giving mean survivals of 842 min (hx546), 810 min (z10), 862 min (z12), and 860 min (z25), compared to 562 min for wild type. All the age-1 alleles were significantly tolerant, compared with wild type (P < 0.001). Two replicates for UV resistance were also completed with mean survivals of 103, 118, 108, and 89 hr, respectively, compared to 72 hr for wild type. One test of hydrogen peroxide resistance has shown that z12 and N2 had a mean survival of 41 hr, while the other age-1 alleles had mean survival of 54 hr (z10), and 62 hr (z25); H2O2 resistance is the only environmental stress that differentiates among the age-1 alleles.
我们使用一种无需影印平板接种的新型筛选方法,分离出了几种新的经甲基磺酸乙酯(EMS)诱变产生的秀丽隐杆线虫长寿突变体。通过不能与age-1(hx546)互补以延长寿命,鉴定出了三个新的age-1等位基因(z10、z12和z25);在25摄氏度时,这些等位基因的寿命在18.9天至25.9天之间,平均寿命延长了46%。回交后,在野生型背景下检测这些等位基因对几种环境胁迫的抗性:热(35摄氏度)、紫外线(UV)(20 J/m2)和过氧化氢(H2O2)(0.5 M)。对每个菌株完成了两次耐热性测试重复实验,野生型的平均存活时间为562分钟,而hx546为842分钟,z10为810分钟,z12为862分钟,z25为860分钟。与野生型相比,所有age-1等位基因都具有显著的耐受性(P < 0.001)。紫外线抗性也完成了两次重复实验,野生型的平均存活时间为72小时,但z10、z12、z25的平均存活时间分别为103小时、118小时、108小时和89小时。一次过氧化氢抗性测试表明,z12和N2的平均存活时间为41小时,而其他age-1等位基因的平均存活时间为54小时(z10)和62小时(z25);过氧化氢抗性是唯一能区分age-1等位基因的环境胁迫。