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一种双筛选方法在果蝇单P因子插入系中鉴定数量性状突变体的高效性。

High efficiency of a double-screening method on single P-element insertion lines to identify quantitative trait mutants in Drosophila melanogaster.

作者信息

Martin Fernando, Kim Min-Su, Gomez-Diaz Carolina, Hovemann Bernhard, Alcorta Esther

机构信息

Depto. Biologia Funcional (Genetica), Fac. Medicina, Universidad de Oviedo, C/ Julian Claveria s/n, 33.006, Oviedo, Spain.

出版信息

Genetica. 2006 Sep-Nov;128(1-3):359-72. doi: 10.1007/s10709-006-7354-z.

Abstract

Enhancer trap P-element insertion has become a common method for generating new mutations in Drosophila melanogaster. When this method is used to isolate mutants for quantitative traits, an appropriate control must be established to define normal and mutant phenotypes. Considering that enhancer-trap lines are generated by crossing several strains, usually with no homogeneous genetic background, no clear control strain can be selected. Previous reports tried to overcome this problem by homogenizing the genetic background of the original lines. However, this is not the most common scenario, especially when functional phenotypes are studied in previously generated lines. Without such caution, is it possible to identify functional mutants among P-element insertion lines? We tested this for olfactory preference, a quantitative trait. Using as control measurement the average phenotype of 30 simultaneously generated P-element insertion lines with preferential reporter-gene expression in olfactory reception organs, we found that 25 of the lines exhibited mutant phenotypes in response to one or several of 5 tested odorants. Additional tests showed that the efficiency of the method for detecting olfactory mutations exceeded 60% even for such a small number of tested odorants. According to these results this approach greatly facilitates the identification of putative abnormal phenotypes, which must be extensively confirmed afterwards.

摘要

增强子捕获P因子插入已成为在黑腹果蝇中产生新突变的常用方法。当使用这种方法分离数量性状的突变体时,必须建立适当的对照来定义正常和突变表型。鉴于增强子捕获系是通过杂交多个品系产生的,通常没有同质的遗传背景,因此无法选择明确的对照品系。先前的报道试图通过使原始品系的遗传背景同质化来克服这个问题。然而,这并非最常见的情况,尤其是在研究先前产生的品系中的功能表型时。如果不注意这一点,是否有可能在P因子插入系中鉴定出功能突变体呢?我们针对嗅觉偏好这一数量性状对此进行了测试。以在嗅觉感受器官中具有优先报告基因表达的30个同时产生的P因子插入系的平均表型作为对照测量,我们发现其中25个品系对5种测试气味剂中的一种或几种表现出突变表型。进一步的测试表明,即使对于如此少量的测试气味剂,该方法检测嗅觉突变的效率也超过了60%。根据这些结果,这种方法极大地促进了推定异常表型的鉴定,不过之后必须进行广泛的确认。

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