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对从二维琼脂糖凝胶中分离并克隆的一些新的果蝇重复DNA序列的分析。

The analysis of some new Drosophila repetitive DNA sequences isolated and cloned from two-dimensional agarose gels.

作者信息

Gilroy T E, Thomas C A

出版信息

Gene. 1983 Jul;23(1):41-51. doi: 10.1016/0378-1119(83)90215-9.

Abstract

Drosophila melanogaster DNA (Dm) was sequentially cleaved by BamHI and EcoRI and separated by two-dimensional gel electrophoresis. Six different prominent bands, which are derived primarily from the cleavage of long sequences that are repeated 20-100 times per genome, were recovered from the gel and cloned in pBR322. Hybridization and restriction analysis of the cloned Dm segments showed that three of these bands are mainly derived from the ribosomal and histone gene repeating units. Segments cloned from the other three bands are not homologous to any known repeating elements that we have tested. They represent long repetitive sequences of moderate multiplicity that appear not to have been hitherto described. These segments have been restriction-mapped and hybridized to cDNA prepared from poly(A)RNA from adult flies. While two minority segments did hybridize to this probe, the majority failed to hybridize. The arrangement of genomic sequences homologous to each plasmid was tested by restriction analysis and Southern hybridization. The results indicate that the repetitive element is largely conserved intact although occupying numerous different positions in the genome. The DNAs from four different strains of D. melanogaster and two of D. simulans produced restriction patterns having some segment lengths in common and some showing clear differences, a fact that indicates that these sequences can move about to occupy different genomic locations in different strains.

摘要

黑腹果蝇DNA(Dm)先用BamHI酶切,再用EcoRI酶切,然后通过二维凝胶电泳进行分离。从凝胶中回收了六个不同的明显条带,这些条带主要来自于每个基因组重复20 - 100次的长序列的切割产物,并克隆到pBR322中。对克隆的Dm片段进行杂交和限制性分析表明,其中三个条带主要来自核糖体和组蛋白基因重复单元。从另外三个条带克隆的片段与我们测试过的任何已知重复元件都不同源。它们代表了中等重复度的长重复序列,似乎迄今尚未被描述过。这些片段已进行限制性图谱分析,并与从成年果蝇的聚腺苷酸RNA制备的cDNA杂交。虽然有两个少数片段确实与该探针杂交,但大多数片段未能杂交。通过限制性分析和Southern杂交测试了与每个质粒同源的基因组序列的排列。结果表明,尽管重复元件在基因组中占据许多不同位置,但它在很大程度上保持完整。来自四种不同黑腹果蝇品系和两种拟果蝇品系的DNA产生的限制性图谱有一些共同的片段长度,也有一些显示出明显差异,这一事实表明这些序列可以移动到不同品系的不同基因组位置。

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