Hartman Phil S, Barry James, Finstad Whitney, Khan Numan, Tanaka Masayuki, Yasuda Kayo, Ishii Naoaki
Department of Biology, Texas Christian University, Fort Worth, TX 76129, United States.
Department of Biology, Texas Christian University, Fort Worth, TX 76129, United States.
Mutat Res. 2014 Aug-Sep;766-767:44-8. doi: 10.1016/j.mrfmmm.2014.05.011. Epub 2014 Jun 10.
Mutagenesis protocols typically call for exposure of late-stage larvae or adults to a mutagen with the intention of inducing mutations in a robust germ line. Instead, ca. 16,000 CB665 [unc-58(e665)] one- to four-cell embryos of the nematode Caenorhabditis elegans were hand selected and exposed to ethyl methanesulfonate (EMS) for 50min. Twenty-one reversion mutants were recovered, of which 17 were intragenic suppressors of the e665 mutation. The mutation frequency was 6.5-fold higher than when CB665 adults were similarly mutagenized, which was predicted given that cell-cycle checkpoints are muted in C. elegans embryos. The mutation spectrum was similar to that obtained after standard EMS mutagenesis.
诱变方案通常要求将晚期幼虫或成虫暴露于诱变剂中,目的是在强大的生殖系中诱导突变。相反,从约16,000个秀丽隐杆线虫的CB665 [unc-58(e665)]一至四细胞胚胎中手工挑选出胚胎,并将其暴露于甲磺酸乙酯(EMS)中50分钟。共获得21个回复突变体,其中17个是e665突变的基因内抑制子。突变频率比用类似方法诱变CB665成虫时高6.5倍,鉴于秀丽隐杆线虫胚胎中的细胞周期检查点被抑制,这一结果在意料之中。突变谱与标准EMS诱变后获得的谱相似。