N-ethyl-N-nitrosourea mutagenesis of a 6- to 11-cM subregion of the Fah-Hbb interval of mouse chromosome 7: Completed testing of 4557 gametes and deletion mapping and complementation analysis of 31 mutations.
N-ethyl-N-nitrosourea mutagenesis of a 6- to 11-cM subregion of the Fah-Hbb interval of mouse chromosome 7: Completed testing of 4557 gametes and deletion mapping and complementation analysis of 31 mutations.
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小鼠7号染色体Fah-Hbb间隔6-11cM亚区的N-乙基-N-亚硝基脲诱变：完成了4557个配子的测试以及31个突变的缺失定位和互补分析。
DOI：
10.1093/genetics/152.1.373
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发表时间：
1999
期刊：
影响因子：
3.3
通讯作者：
Carpenter,DA
中科院分区：
文献类型：
--
作者：
Rinchik,EM;Carpenter,DA
小鼠染色体(Chr)7围绕白化(Tyr；c)基因座的间隔，对应于6到11 cM的Tyr长缺失，已成为化学超诱变剂N-乙基-N-亚硝胺(ENU)大规模诱变筛选的目标。来自ENU处理雄鼠的诱变基因组中的一段Chr 7被半合，与长缺失相反，以识别和恢复映射在白化删除复合体内的新的隐性突变。对6000多个家系进行了分析，其中4557个家系进行了完整的突变测试，确定了致死性和肉眼可见的表型。通过两两反式互补杂交，鉴定出31个非聚集性突变，并将其分配到10个互补组。利用广泛的辐射诱导的Tyr缺失系列的缺失作图分析，将这些互补组中的每一个所定义的座位放置到Tyr区域缺失图谱的指定区间中，这有助于在该区域的物理和转录图谱上识别每个座位。这些突变确定了7个新的基因座，并在先前定义的3个基因座上提供了ENU诱导的新等位基因。有趣的是，通过对白化病纯合子或部分互补缺失的分析，没有发现重述三种表型的突变。根据我们对这一筛选的经验，我们讨论了在考虑将化学诱变筛选应用于小鼠基因组的百万碱基区时所关注的一些问题(例如，基因座突变、饱和失败、要筛选的配子数量、等位基因序列)。
An interval of mouse chromosome (Chr) 7 surrounding the albino (Tyr; c) locus, and corresponding to a long 6- to 11-cM Tyr deletion, has been the target of a large-scale mutagenesis screen with the chemical supermutagen N-ethyl-N-nitrosourea (ENU). A segment of Chr 7, from a mutagenized genome bred from ENU-treated males, was made hemizygous opposite the long deletion for recognition and recovery of new recessive mutations that map within the albino deletion complex. Over 6000 pedigrees were analyzed, and 4557 of these were completely tested for mutations specifying both lethal and gross visible phenotypes. Thirty-one nonclustered mutations were identified and assigned to 10 complementation groups by pairwise trans-complementation crosses. Deletion-mapping analyses, using the extensive series of radiation-induced Tyr deletions, placed the loci defined by each of these complementation groups into defined intervals of the Tyr-region deletion map, which facilitates the identification of each locus on physical and transcription maps of the region. These mutations identified seven new loci and provided new ENU-induced alleles at three previously defined loci. Interestingly, no mutations were recovered that recapitulated three phenotypes defined by analysis of homozygous or partially complementing albino deletions. On the basis of our experience with this screen, we discuss a number of issues (e.g., locus mutability, failure to saturate, number of gametes to screen, allelic series) of concern when application of chemical mutagenesis screens to megabase regions of the mouse genome is considered.