Optimization of ENU mutagenesis of Caenorhabditis elegans
Optimization of ENU mutagenesis of Caenorhabditis elegans
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DOI：
10.1016/s1383-5718(01)00198-x
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发表时间：
2001-08-22
影响因子：
1.9
通讯作者：
Dorman, S
中科院分区：
文献类型：
--
作者：
De Stasio, EA;Dorman, S
秀丽隐杆线虫的化学突变主要依赖于EMS来产生错义突变。EMS诱变的缺点是分子损伤主要是G/C -> A/T转换。ENU已被证明产生不同的突变谱，但其对C. elegans使得它很难成为诱变剂。我们在这里描述的方法，最大限度地减少ENU毒性在C。优美的方法包括在无水乙醇中制备ENU储备液，并将储备液在-20 ℃下储存不超过2周。为了保持诱变动物的合理窝数，诱变溶液不应超过1.0 mM ENU。我们提供的数据表明ENU在水溶液中降解或改变为毒性更大的产物，但在溶剂如无水乙醇中则较少。(C)出版社：Elsevier Science B. V.
Chemical mutagenesis of Caenorhabditis elegans has relied primarily on EMS to produce missense mutations. The drawback of EMS mutagenesis is that the molecular lesions are primarily G/C --> A/T transitions. ENU has been shown to produce a different spectrum of mutations, but its greater toxicity to C. elegans makes it a difficult mutagen to use. We describe here methods for minimizing ENU toxicity in C. elegans. Methods include preparing ENU stocks in absolute ethanol and storing stock solutions for not more than 2 weeks at -20 degreesC. To maintain reasonable brood sizes of mutagenized animals, mutagenic solutions should not exceed 1.0 mM ENU. We provide data which suggest ENU is degraded or altered to more toxic products in aqueous solution, but less so in solvents such as absolute ethanol. (C) 2001 Published by Elsevier Science B.V.