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[黑腹果蝇核糖体RNA中28S基因插入序列的克隆与研究]

[Cloning and study of inserted sequences of gene 28S in Drosophila melanogaster ribosomal RNA].

作者信息

Kolchinskiĭ A M, Vashakidze R P, Evgen'ev M B, Mirzabekov A D

出版信息

Mol Biol (Mosk). 1982 Mar-Apr;16(2):302-14.

PMID:6280035
Abstract

Cloning of fragments of ribosomal genes containing insertions in the 28S RNA gene has been reported earlier. Subcloning of DNA fragments corresponding to insertion sequences and their hybridization with DNA, RNA and polytene chromosomes from different flies is described. Type 1 insertions (containing BamI sites) are highly heterogeneous in length and sequence even in homozygotes. Type 2 insertions (with EcoRI sites) are rather homogeneous. Two types of insertions are represented in the D. melanogaster genome by 50 and 30 copies, respectively. Restriction fragments with insertions significantly differ in DNA from embryos and larvae. D. simulans and D. virilis also contain the sequences of both types of insertions, though in fewer number of copies. Type 1 insertions seem to be poorly transcribed, and type 2 insertions are not transcribed at all. Among 2000 recombinant clones screened a number of DI plasmids hybridizing to isolated insertions were obtained. Six of them were mapped with restriction endonucleases and hybridized with insertion fragments. rRNA and polytene chromosomes. All of these DI plasmids hybridize with the nucleoli, one with the chromocenter and one with the 79F 3L site. In LI9, not coding for rRNA, the sequences, corresponding to two types on insertions are located only a few kilobases apart. D17a does not encode for rRNA, but hybridizes in situ only with the nucleoli.

摘要

此前已有报道克隆出在28S RNA基因中含有插入片段的核糖体基因片段。本文描述了与插入序列相对应的DNA片段的亚克隆及其与来自不同果蝇的DNA、RNA和多线染色体的杂交情况。1型插入片段(含有BamI位点)即使在纯合子中长度和序列也高度异质。2型插入片段(含有EcoRI位点)则较为均一。在黑腹果蝇基因组中,两种类型的插入片段分别有50个和30个拷贝。含有插入片段的限制性片段在胚胎和幼虫的DNA中有显著差异。拟果蝇和粗壮果蝇也含有这两种类型插入片段的序列,不过拷贝数较少。1型插入片段似乎转录不良,2型插入片段则根本不转录。在筛选的2000个重组克隆中,获得了一些与分离出的插入片段杂交的DI质粒。其中6个用限制性内切酶进行了图谱分析,并与插入片段、rRNA和多线染色体杂交。所有这些DI质粒都与核仁杂交,一个与染色中心杂交,一个与79F 3L位点杂交。在不编码rRNA的LI9中,与两种类型插入片段相对应的序列仅相隔几千个碱基。D17a不编码rRNA,但仅在原位与核仁杂交。

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