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两栖类灯刷染色体标志性环的DNA显微切割与克隆

Microdissection and cloning of DNA from landmark loops of amphibian lampbrush chromosomes.

作者信息

Penrad-Mobayed M, Sourrouille P, Bonnanfant-Jaïs M L, N'Da E, Edström J E, Angelier N

机构信息

Centre de Biologie Cellulaire, UPR 3101, Ivry-sur-Seine, France.

出版信息

Chromosoma. 1991 Dec;101(3):180-8. doi: 10.1007/BF00355367.

Abstract

Microdissection of the "globular" and "granular" landmark loops of Pleurodeles lampbrush chromosomes and subsequent cloning of their DNA yielded several recombinant clones. The 6.6-kb insert of one of them was subcloned and the 600 bp of one subclone was characterized by Southern and slot hybridizations as well as by sequencing. This sequence, designated p130B, was shown to belong to a class of moderately repetitive DNA. RNA expression of this sequence was investigated by in situ hybridization of p130B to the nascent transcripts of lateral loops. Results showed that: (1) the same transcripts were not always found in matrices of landmarks exhibiting the same morphological features; (2) the same transcripts were expressed in loops of different morphological types. Based on these results we suggest that even if there is a morphological similarity of landmark loops, this does not reflect total similarity of their transcripts.

摘要

对美西螈灯刷染色体的“球状”和“颗粒状”标志性环进行显微切割,随后对其DNA进行克隆,获得了几个重组克隆。其中一个克隆的6.6kb插入片段被亚克隆,一个亚克隆的600bp通过Southern杂交、狭缝杂交以及测序进行了表征。这个序列被命名为p130B,属于中度重复DNA类别。通过将p130B与侧环的新生转录本进行原位杂交,研究了该序列的RNA表达。结果表明:(1)在具有相同形态特征的标志性结构的基质中,并非总能发现相同的转录本;(2)相同的转录本在不同形态类型的环中表达。基于这些结果,我们认为,即使标志性环在形态上相似,这也并不反映它们转录本的完全相似性。

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