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

机译:高效率的双重筛选方法在单个P元素插入线上鉴定果蝇的数量性状突变体

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摘要

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.
机译:增强子陷阱P元素的插入已成为在果蝇中产生新突变的常用方法。当此方法用于分离突变体的数量性状时,必须建立适当的对照来定义正常和突变表型。考虑到增强子-捕获系是通过交叉多个菌株而产生的,通常没有同质的遗传背景,因此无法选择明确的对照菌株。先前的报道试图通过使原始品系的遗传背景均质来克服这个问题。但是,这不是最常见的情况,尤其是在先前生成的品系中研究功能表型时。如果没有这样的谨慎,是否有可能在P元素插入系中鉴定功能性突变体?我们针对嗅觉偏好(一种定量特征)进行了测试。使用30条同时产生的P元素插入序列在嗅觉接收器官中具有优先报告基因表达的平均表型作为对照测量,我们发现其中的25条系表现出对5种测试气味剂中的一种或几种有响应的突变表型。附加测试表明,即使对于如此少量的测试气味剂,检测嗅觉突变的方法的效率也超过60%。根据这些结果,该方法极大地促进了推定的异常表型的鉴定,此后必须进行广泛的确认。

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