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高重复DNA与有冠蝾螈染色体的原位杂交。

In situ hybridization of highly repetitive DNA to chromosomes of Triturus cristatus.

作者信息

Macgregor H C

出版信息

Chromosoma. 1979 Feb 13;71(1):57-64. doi: 10.1007/BF00426366.

Abstract

Highly repetitive DNA of C0t 0--0.2 was purified from whole DNA of Triturus cristatus carnifex, labelled by nick translation, and in situ hybridized to RNA transcripts on the loops of lampbrush chromosomes and to the DNA of mitotic chromosomes from intestinal epithelium from the same species. The labelled DNA bound to 20--30 loops on the long arms of lampbrush bivalent 1, a pair of loops near the centromere on bivalent 10, and a number of other loops most of which were localized in pericentric regions. In mitotic preparations the same labelled DNA bound to the heteromorphic regions of the long arms of both chromosomes 1, and to the centromeric regions of all chromosomes. Centromeric labelling was light on chromosomes 4 and particularly clear on the 3 shortest chromosomes of the set. The heavy labelling of the heteromorphic arms of chromosome 1 is discussed in relation to several other peculiar properties of these arms, including their extraordinary lengths, their Giemsa banding patterns, and the absence of meiotic crossing over. It is suggested that insofar as the results with DNA/DNA hybridization and mitotic chromosomes match those obtained with the DNA/RNA-transcript hybridization and lampbrush chromosomes, confidence in the latter technique may be increased accordingly.

摘要

从真螈(Triturus cristatus carnifex)的全基因组DNA中纯化出C0t值为0--0.2的高度重复DNA,通过缺口平移法进行标记,并与灯刷染色体环上的RNA转录本以及来自同一物种肠上皮细胞有丝分裂染色体的DNA进行原位杂交。标记的DNA与灯刷二价体1长臂上的20--30个环、二价体10着丝粒附近的一对环以及许多其他环结合,其中大多数位于着丝粒周围区域。在有丝分裂标本中,相同的标记DNA与两条1号染色体长臂的异形区域以及所有染色体的着丝粒区域结合。4号染色体上的着丝粒标记较浅,而在该染色体组中最短的3条染色体上则特别明显。结合染色体1异形臂的其他几个特殊特性,包括其超长的长度、吉姆萨带型以及减数分裂过程中不存在交叉互换现象,对染色体1异形臂的强标记现象进行了讨论。结果表明,鉴于DNA/DNA杂交和有丝分裂染色体的结果与DNA/RNA转录本杂交和灯刷染色体的结果相匹配,因此可以相应地增强对后一种技术的信心。

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