Molecular characterization of ENU mouse mutagenesis and archives
Molecular characterization of ENU mouse mutagenesis and archives
复制标题
DOI：
10.1016/j.bbrc.2005.08.134
复制
发表时间：
2005-10-21
影响因子：
3.1
通讯作者：
Gondo, Y
中科院分区：
文献类型：
--
作者：
Sakuraba, Y;Sezutsu, H;Gondo, Y
ENU大规模诱变小鼠为功能基因组学研究提供了前沿遗传资源。冷冻精子档案的ENU诱变的第1代（G1）小鼠也可以提供一个“突变小鼠库”，使我们能够进行反向遗传学在任何特定的目标基因。我们已经存档了9224只G1小鼠的冷冻精子和基因组DNA。通过全基因组筛选63个靶基因座，覆盖了小鼠基因组的197 Mbp的总和，总共148个ENU诱导的突变已被直接确定。突变位点的初步鉴定采用温度梯度毛细管电泳法，然后直接测序。分子生物学鉴定结果表明，所有鉴定的突变均为点突变，除少数冗余突变外，大部分为独立事件。本研究中的碱基取代谱不同于基于表型的诱变。基于ENU的小鼠基因驱动突变现在变得可行且实用。(c)2005年爱思唯尔公司All rights reserved.
The large-scale mouse mutagenesis with ENU has provided forward-genetic resources for functional genomics. The frozen sperm archive of ENU-mutagenized generation-1 (G1) mice could also provide a "mutant mouse library" that allows us to conduct reverse genetics in any particular target genes. We have archived frozen sperm as well as genomic DNA from 9224 G1 mice. By genome-wide screening of 63 target loci covering a sum of 197 Mbp of the mouse genome, a total of 148 ENU-induced mutations have been directly identified. The sites of mutations were primarily identified by temperature gradient capillary electrophoresis method followed by direct sequencing. The molecular characterization revealed that all the identified mutations were point mutations and mostly independent events except a few cases of redundant mutations. The base-substitution spectra in this study were different from those of the phenotype-based mutagenesis. The ENU-based gene-driven mutagenesis in the mouse now becomes feasible and practical. (c) 2005 Elsevier Inc. All rights reserved.