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通过缺口平移和荧光原位杂交对豌豆蚜(同翅目,蚜科)中一个高度重复的EcoRI DNA片段进行染色体定位

Chromosomal localization of a highly repeated EcoRI DNA fragment in Megoura viciae (Homoptera, Aphididae) by nick translation and fluorescence in situ hybridization.

作者信息

Bizzaro D, Manicardi G C, Bianchi U

机构信息

Dipartimento di Biologia Animale, Università di Modena, Italy.

出版信息

Chromosome Res. 1996 Aug;4(5):392-6. doi: 10.1007/BF02257275.

DOI:10.1007/BF02257275
PMID:8871828
Abstract

To investigate the genome of the aphid Megoura viciae at molecular level, we have studied total DNA by agarose gel electrophoresis after cleavage with different restriction endonucleases. EcoRI digestion produced a highly repeated DNA fragment, about 600 pb long. The contribution of this EcoRI element to the total genome of M. viciae was estimated at about 6% by means of densitometric scanning of agarose gel photographs. The chromosomal localization of this fragment, investigated by fluorescent in situ hybridization (FISH), constantly showed one large and two narrower fluorescent bands located on the X chromosome, all corresponding to C-positive heterochromatic areas. These results are in full accordance with the data obtained by in situ nick translation experiments carried out after EcoRI digestion, and clearly demonstrate that a substantial amount of M. viciae heterochromatin consists of EcoRI fragments which are mainly located on the X chromosome. Using the EcoRI restriction fragment as a molecular probe may be a practical tool for the investigation of taxonomic and evolutionary relationships in this group of insects.

摘要

为了在分子水平上研究豌豆蚜(Megoura viciae)的基因组,我们在用不同的限制性内切酶切割后,通过琼脂糖凝胶电泳对总DNA进行了研究。EcoRI酶切产生了一个高度重复的DNA片段,长度约为600 pb。通过对琼脂糖凝胶照片进行密度扫描,估计这个EcoRI元件对豌豆蚜总基因组的贡献约为6%。通过荧光原位杂交(FISH)研究该片段的染色体定位,结果始终显示在X染色体上有一条大的和两条较窄的荧光带,均对应于C阳性异染色质区域。这些结果与EcoRI酶切后进行的原位缺口平移实验所获得的数据完全一致,并且清楚地表明,大量的豌豆蚜异染色质由主要位于X染色体上的EcoRI片段组成。使用EcoRI限制性片段作为分子探针可能是研究这类昆虫分类学和进化关系的一种实用工具。

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