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首页> 外文期刊>Journal of Fish Biology >Transgene-mediated skeletal phenotypic variation in zebrafish
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Transgene-mediated skeletal phenotypic variation in zebrafish

机译：转基因介导斑马鱼的骨骼表型变异

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When considering relationships between genotype and phenotype we frequently ignore the fact that the genome of a typical animal, notably including that of a fish and a human, harbours a huge amount of foreign DNA. Such DNA, in the form of transposable elements, can affect genome function in a major way, and transgene biology needs to be included in our understanding of the genome. Here we examine an unexpected phenotypic effect of the chromosomally integrated transgene fli1a-F-hsp70l:Gal4VP16 that serves as a model for transgene function generally. We examine larval fras1 mutant zebrafish (Danio rerio). Gal4VP16 is a potent transcriptional activator that is already well known for toxicity and mediating unusual transcriptional effects. In the presence of the transgene, phenotypes in the neural crest-derived craniofacial skeleton, notably fusions and shape changes associated with loss of function fras1 mutations, are made more severe, as we quantify by scoring phenotypic penetrance, the fraction of mutants expressing the trait. A very interesting feature is that the enhancements are highly specific for fras1 mutant phenotypes, occurring in the apparent absence of more widespread changes. Except for the features due to the fras1 mutation, the transgene-bearing larvae appear generally healthy and to be developing normally. The transgene behaves as a genetic partial dominant: a single copy is sufficient for the enhancements, yet, for some traits, two copies may exert a stronger effect. We made new strains bearing independent insertions of the fli1a-F-hsp70l:Gal4VP16 transgene in new locations in the genome, and observed increased severities of the same phenotypes as observed for the original insertion. This finding suggests that sequences within the transgene, for example Gal4VP16, are responsible for the enhancements, rather than the effect on neighbouring host sequences (such as an insertional mutation). The specificity and biological action underlying the traits are subjects of considerable interest for further investigation, as we discuss. Our findings show that work with transgenes needs to be undertaken with caution and attention to detail.
机译：在考虑基因型和表型之间的关系时，我们经常忽略一个事实，即典型动物的基因组，尤其是鱼类和人类的基因组，含有大量外源DNA。这种以转座因子形式存在的DNA可以在很大程度上影响基因组功能，我们对基因组的理解需要包括转基因生物学。在这里，我们研究了染色体整合的转基因fli1a-F-hsp70l:Gal4VP16的一种意想不到的表型效应，该基因通常作为转基因功能的模型。我们检测了fras1突变斑马鱼（Danio rerio）的幼虫。Gal4VP16是一种有效的转录激活剂，其毒性和介导不寻常的转录效应已经众所周知。在存在转基因的情况下，神经嵴衍生的颅面骨骼中的表型，尤其是与功能丧失fras1突变相关的融合和形状变化，变得更加严重，因为我们通过评分表型外显率来量化，外显率是指表达该特征的突变体的比例。一个非常有趣的特征是，增强对fras1突变表型具有高度特异性，发生在明显没有更广泛变化的情况下。除了fras1突变引起的特征外，携带转基因的幼虫似乎总体健康，发育正常。转基因表现为遗传部分显性：一个拷贝就足以增强，然而，对于某些性状，两个拷贝可能会产生更强的效果。我们在基因组的新位置制造了携带fli1a-F-hsp70l:Gal4VP16转基因独立插入的新菌株，并观察到与原始插入相同表型的严重性增加。这一发现表明，转基因内的序列，例如Gal4VP16，是增强的原因，而不是对邻近宿主序列的影响（例如插入突变）。正如我们所讨论的，这些特征背后的特异性和生物学作用是值得进一步研究的主题。我们的发现表明，转基因工作需要谨慎进行，并注意细节。

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