Rosenbluth R E, Cuddeford C, Baillie D L
Genetics. 1985 Mar;109(3):493-511. doi: 10.1093/genetics/109.3.493.
We previously established a gamma-ray dose-response curve for recessive lethal events (lethals) captured over the eT1 balancer. In this paper we analyze the nature of lethal events produced, with a frequency of 0.04 per eT1 region, at a dose of 1500 r. To do so, we developed a protocol that, in the absence of cytogenetics, allows balanced lethals to be analyzed for associated chromosomal rearrangements. A set of 35 lethal strains was chosen for the analysis. Although the dosage was relatively low, a large number of multiple-break events were observed. The fraction of lethals associated with rearrangements was found to be 0.76. Currently most X- and gamma-ray dosages used for mutagenesis in C. elegans are 6000-8000 r. From our data we conservatively estimated that 43% of rearrangements induced with 8000 r would be accompanied by additional chromosome breaks in the genome. With 1500 r the value was 5%. The 35 lethals studied were derived from 875 screened F1's. Among these lethals there were (1) at least two unc-36 duplications, (2) at least four translocations, (3) at least six deficiencies of chromosome V (these delete about 90% of the unc-60 to unc-42 region) and (4) several unanalyzed rearrangements. Thus, it is possible to recover desired rearrangements at reasonable rates with a dose of only 1500 r. We suggest that the levels of ionizing radiation employed in most published C. elegans studies are excessive and efforts should be made to use reduced levels in the future.
我们之前建立了一条γ射线剂量-反应曲线,用于分析在eT1平衡染色体上捕获的隐性致死事件(致死突变)。在本文中,我们分析了在1500伦琴剂量下产生的致死事件的性质,其频率为每eT1区域0.04。为此,我们开发了一种方案,在没有细胞遗传学的情况下,该方案允许对平衡致死突变进行相关染色体重排分析。选择了一组35个致死菌株进行分析。尽管剂量相对较低,但观察到大量的多重断裂事件。发现与重排相关的致死突变比例为0.76。目前,秀丽隐杆线虫诱变中使用的大多数X射线和γ射线剂量为6000 - 8000伦琴。根据我们的数据,我们保守估计,8000伦琴诱导的重排中有43%会伴随着基因组中额外的染色体断裂。在1500伦琴剂量下,这一数值为5%。所研究的35个致死突变源自875个筛选的F1代。在这些致死突变中,有(1)至少两个unc-36重复,(2)至少四个易位,(3)至少六个五号染色体缺失(这些缺失大约90%的unc-60到unc-42区域),以及(4)几个未分析的重排。因此,仅用1500伦琴的剂量就有可能以合理的速率获得所需的重排。我们认为,大多数已发表的秀丽隐杆线虫研究中使用的电离辐射水平过高,未来应努力使用更低的水平。