Sequence-based analysis of mutagenized mice
Sequence-based analysis of mutagenized mice
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DOI：
10.1007/s003350010113
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发表时间：
2000-07-01
期刊：
影响因子：
2.5
通讯作者：
Beier, DR
中科院分区：
文献类型：
--
作者：
Beier, DR
用二亚硝基脲 (ENU) 治疗小鼠是诱变精原细胞的有效方法，这种治疗方法已在各种显性和隐性突变筛选中被证明有效。然而，该技术的一个重要问题是，诱变效率最常通过使用特定基因座测试经验确定每个基因座突变频率来评估，这是昂贵、耗时且逻辑上困难的。为了更直接、更有效地解决这个问题，可以利用基于 PCR 的突变检测方法来表征诱变小鼠的后代。由于这种分析可以在单代育种后完成，因此它可作为评估诱变剂处理效率的快速方法。此外，很容易想象该策略可以应用于以系统方式一般确定基因功能。理论考虑和实证分析表明，分次剂量治疗方案的每个碱基突变频率约为每 10(5) bp 1 个序列变化。
Treating mice with erhylnitrosourea (ENU) is an efficient means for mutagenizing spermatogonial cells, and this treatment has been proven effective in a variety of screens for both dominant and recessive mutations. However, a significant problem for this technology is that the efficiency of mutagenesis is assessed most often by the empiric determination of a per-locus mutation frequency by using the specific locus test, which is expensive, time-consuming, and logistically difficult. To approach this question more directly and more efficiently, one can utilize methods of PCR-based mutation detection for the characterization of progeny of mutagenized mice. Since this analysis can be done after a single generation of breeding, it is useful as a rapid means for the assessment of the efficiency of mutagen treatment. Furthermore, it is readily imaginable that this strategy can be applied for the general determination of gene function in a systematic manner. Theoretical considerations and empirical analysis suggest that the per-base mutation frequency for a fractionated-dose treatment protocol is on the order of 1 sequence change per 10(5) bp.