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直接测定基因型和极端温度对黑腹果蝇长期突变积累品系中roo转座的影响。

Direct determination of the effects of genotype and extreme temperature on the transposition of roo in long-term mutation accumulation lines of Drosophila melanogaster.

作者信息

Vázquez J Fernando, Albornoz Jesús, Domínguez Ana

机构信息

Departamento de Biología Funcional, Area de Genética, Facultad de Medicina, Universidad de Oviedo, Julián Clavería s/n, 33071 Oviedo, Spain.

出版信息

Mol Genet Genomics. 2007 Dec;278(6):653-64. doi: 10.1007/s00438-007-0282-5. Epub 2007 Aug 25.

DOI:10.1007/s00438-007-0282-5
PMID:17721789
Abstract

Transposable elements (TEs) are mobile repetitive DNA sequences that constitute a structurally dynamic component of genomes. In order to understand the dynamics of TEs it is necessary to have information about the control of transposition and its dependence of environmental factors. After a great deal of previous work on transposition conducted on long-term mutation accumulation (MA) lines of Drosophila melanogaster started in 1987, only roo out of 16 families was found active in this genotype. Here we test the effect of the modification of the genetic background by introducing a Cy chromosome, and the effect of extreme temperature (28 degrees C) on the transposition rate of roo. Thermal stress did not affect the transposition rate, whereas the presence of a Cy chromosome in heterozygosis lowered it. There was an excess of insertions in the X chromosome, with respect to autosomes, and in the proximal and distal regions of chromosome arms that can be interpreted as target site preference. One of the control lines became highly unstable with mean insertion and excision rates of 3.0 x 10(-3) and 8.5 x 10(-4), respectively. Instability arose spontaneously during generations of mutation accumulation, and can be attributed to "de novo" mutation. Transposition in the unstable line could be directly studied on the progeny of individual males and females, from where we deduced that transposition occurs mainly, if not exclusively, in males, with a rate of 1.125 insertions per gamete. In situ hybridization with an LTR probe showed that most excisions (12 out of 14) were precise. Our data show the prominent role of genotype in transposition control and can explain rapid turnovers in the genome without increasing the number of copies.

摘要

转座元件(TEs)是可移动的重复DNA序列,构成了基因组中结构动态变化的组成部分。为了理解转座元件的动态变化,有必要了解转座控制及其对环境因素的依赖性。自1987年开始对黑腹果蝇的长期突变积累(MA)品系进行了大量关于转座的前期研究后,在16个家族中仅发现roo在该基因型中具有活性。在此,我们测试了引入Cy染色体对遗传背景修饰的影响,以及极端温度(28摄氏度)对roo转座率的影响。热应激并未影响转座率,而异合子状态下Cy染色体的存在则降低了转座率。相对于常染色体,X染色体以及染色体臂的近端和远端区域存在过多的插入,这可解释为靶位点偏好。其中一个对照品系变得高度不稳定,平均插入率和切除率分别为3.0×10⁻³和8.5×10⁻⁴。不稳定性在突变积累的世代中自发出现,可归因于“从头”突变。不稳定品系中的转座可直接在单个雄性和雌性的后代中进行研究,由此我们推断转座主要(如果不是唯一的话)发生在雄性中,每个配子的插入率为1.125次。用LTR探针进行原位杂交显示,大多数切除(14次中有12次)是精确的。我们的数据表明基因型在转座控制中起重要作用,并且可以解释基因组中无需增加拷贝数的快速周转。

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