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Mos1介导的秀丽隐杆线虫插入诱变

Mos1-mediated insertional mutagenesis in Caenorhabditis elegans.

作者信息

Boulin Thomas, Bessereau Jean-Louis

机构信息

ENS, 46 rue d'Ulm, F-75005 Paris, France.

出版信息

Nat Protoc. 2007;2(5):1276-87. doi: 10.1038/nprot.2007.192.

Abstract

We describe a protocol for mutating genes in the nematode Caenorhabditis elegans using the Mos1 transposon of Drosophila mauritiana. Mutated genes containing a Mos1 insertion are molecularly tagged by this heterologous transposable element. Mos1 insertions can therefore be identified in as little as 3 weeks using only basic molecular biology techniques. Mutagenic efficiency of Mos1 is tenfold lower than classical chemical mutagens. However, the ease and speed with which mutagenic insertions can be mapped compares favorably with the vast amount of work involved in classical genetic mapping. Therefore, Mos1 could be the tool of choice when screening procedures are efficient. In addition, Mos1 mutagenesis can greatly simplify the mapping of mutations that exhibit low penetrance, subtle or synthetic phenotypes. The recent development of targeted engineering of C. elegans loci carrying Mos1 insertions further increases the attractiveness of Mos1-mediated mutagenesis.

摘要

我们描述了一种使用毛里求斯果蝇的Mos1转座子对线虫秀丽隐杆线虫中的基因进行突变的方案。含有Mos1插入的突变基因被这种异源转座元件进行分子标记。因此,仅使用基本的分子生物学技术,在短短3周内就可以鉴定出Mos1插入。Mos1的诱变效率比传统化学诱变剂低十倍。然而,诱变插入的定位简便性和速度与经典遗传定位所涉及的大量工作相比具有优势。因此,当筛选程序高效时,Mos1可能是首选工具。此外,Mos1诱变可以极大地简化对表现出低外显率、细微或合成表型的突变的定位。携带Mos1插入的秀丽隐杆线虫基因座的靶向工程的最新进展进一步增加了Mos1介导的诱变的吸引力。

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