Sensitivity of Allelic Divergence to Genomic Position: Lessons from the Drosophila tan Gene.
Sensitivity of Allelic Divergence to Genomic Position: Lessons from the Drosophila tan Gene.
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DOI：
10.1534/g3.116.032029
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发表时间：
2016-09-08
期刊：
影响因子：
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通讯作者：
Wittkopp PJ
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文献类型：
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作者：
John AV;Sramkoski LL;Walker EA;Cooley AM;Wittkopp PJ
为了确定物种内和物种之间表型变化的遗传变异，研究人员经常利用转基因动物来比较不同遗传背景中等位基因的功能。在果蝇中，由ΦC31整合酶介导的靶向整合允许在相同基因组位置比较替代等位基因的活性。通过对每个转基因使用相同的插入位点，位置效应通常被认为是受控的，因为两个等位基因被相同的基因组背景包围。在这里，我们通过比较两种果蝇的tan等位基因的活性来验证这一假设。americana和D. novamexicana，在D.黑腹菌我们发现，这些等位基因的相对影响不同的插入位点，在两个网站之间观察到的活动没有差异。其中一个位点简单地沉默了两个转基因，但另一个位点允许两个等位基因的表达，这足以拯救突变表型，但未能揭示两个等位基因之间的功能差异。这些结果表明，在比较转基因的活性时，应该使用一个以上的插入位点，因为如果不这样做，可能会导致等位基因之间的功能差异未被检测到。
To identify genetic variants underlying changes in phenotypes within and between species, researchers often utilize transgenic animals to compare the function of alleles in different genetic backgrounds. In Drosophila, targeted integration mediated by the ΦC31 integrase allows activity of alternative alleles to be compared at the same genomic location. By using the same insertion site for each transgene, position effects are generally assumed to be controlled for because both alleles are surrounded by the same genomic context. Here, we test this assumption by comparing the activity of tan alleles from two Drosophila species, D. americana and D. novamexicana, at five different genomic locations in D. melanogaster. We found that the relative effects of these alleles varied among insertion sites, with no difference in activity observed between them at two sites. One of these sites simply silenced both transgenes, but the other allowed expression of both alleles that was sufficient to rescue a mutant phenotype yet failed to reveal the functional differences between the two alleles. These results suggest that more than one insertion site should be used when comparing the activity of transgenes because failing to do so could cause functional differences between alleles to go undetected.