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Exploring Supernumeraries - A New Marker for Screening of B-Chromosomes Presence in the Yellow Necked Mouse Apodemus flavicollis

机译:探索数字-筛选黄色颈鼠黄饰线虫B染色体存在的新标记

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摘要

Since the density of simple sequence repeats (SSRs) may vary between different chromosomes of the same species in eukaryotic genomes, we screened SSRs of the whole genome of the yellow necked mouse, Apodemus flavicollis, in order to reveal SSR profiles specific for animals carrying B chromosomes. We found that the 2200 bp band was amplified by primer (CAG)4AC to a highly increased level in samples with B chromosomes. This quantitative difference (B-marker) between animals with (+B) and without (0B) B chromosomes was used to screen 20 populations (387 animals). The presence/absence of Bs was confirmed in 96.5% of 342 non mosaic individuals, which recommends this method for noninvasive B-presence detection. A group of 45 animals with mosaic and micro B (μB) karyotypes was considered separately and showed 55.6% of overall congruence between karyotyping and molecular screening results. Relative quantification by qPCR of two different targeted sequences from B-marker indicated that these B-specific fragments are multiplied on B chromosomes. It also confirms our assumption that different types of Bs with variable molecular composition may exist in the same individual and between individuals of this species. Our results substantiate the origin of Bs from the standard chromosomal complement. The B-marker showed 98% sequence identity with the serine/threonine protein kinase VRK1 gene, similarly to findings reported for Bs from phylogenetically highly distant mammalian species. Evolutionarily conserved protein-coding genes found in Bs, including this one in A. flavicollis, could suggest a common evolutionary pathway.
机译:由于真核生物基因组中同一物种的不同染色体之间的简单序列重复(SSR)密度可能不同,因此我们筛选了黄颈小鼠Apodemus flavicollis整个基因组的SSR,以揭示特异于携带B的动物的SSR概况染色体。我们发现,在带有B染色体的样本中,引物(CAG)4AC将2200 bp的条带扩增到高度增加的水平。具有(+ B)B染色体和不具有(0B)B染色体的动物之间的数量差异(B标记)用于筛选20个种群(387只动物)。在342个非镶嵌个体中,有96.5%的人确认了Bs的存在/不存在,这建议将该方法用于无创性B存在检测。一组45只具有镶嵌和微型B(μB)核型的动物被单独考虑,它们在核型分析和分子筛选结果之间的总体一致性为55.6%。通过qPCR对来自B标记的两个不同靶向序列的相对定量表明,这些B特异性片段在B染色体上倍增。这也证实了我们的假设,即在同一个体中以及该物种的个体之间可能存在具有可变分子组成的不同类型的Bs。我们的结果证实了标准染色体补体中Bs的起源。 B标记显示与丝氨酸/苏氨酸蛋白激酶VRK1基因有98%的序列同一性,与从系统发育高度远距离的哺乳动物物种获得的Bs报道的发现相似。在Bs中发现进化保守的蛋白质编码基因,包括在黄曲霉中的这一基因,可能暗示了一条共同的进化途径。

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