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湖红点鲑(Salvelinus namaycush)高度重复DNA的分子特征及细胞遗传学分析

Molecular characterization and cytogenetic analysis of highly repeated DNAs of lake trout, Salvelinus namaycush.

作者信息

Reed K M, Phillips R B

机构信息

Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI 53201, USA.

出版信息

Chromosoma. 1995 Dec;104(4):242-51. doi: 10.1007/BF00352255.

Abstract

The chromosomes of lake trout (Salvelinus namaycush) contain a considerable amount of heterochromatin located at the centromeres and/or telomeres of several chromosomes, including a sex-specific block located distally on the X chromosome. In order to investigate further the repetitive DNAs of lake trout, genomic DNA from a female was size fractionated (<600 bp) with the restriction endonuclease AluI and fragments were cloned into the bacteriophage M13. A total of 42 clones were isolated. Relative copy number of individual inserts within the lake trout genome was estimated by Southern analysis. Twelve clones were determined to be highly repetitive and were chosen for further investigation. Inserts of these clones contained sequences similar to the AluI/RsaI, EcoRI/DraI, DraI/BstEII, and MboI/BglII families reported from Arctic char (Salvelinus alpinus). The chromosomal location of several of these fragments was determined in lake trout by fluorescence in situ hybridization (FISH). Two related AluI/RsaI sequences (Type A, approximately 140 bp, and Type B,approximately 120 bp) showed differential hybridization. Type A hybridized to the centromeres of all metacentric as well as several acrocentric chromosomes. Type B hybridized to the centromeres of most acrocentric chromosomes. A sequence with homology to the EcoRI/DraI family hybridized to the centromeres of several acrocentric chromosomes. Sequences with partial similarity to the DraI/BstEII family hybridized to the major rDNA sites (nucleolar organizer regions, NORs) and several minor telomeric sites. The interstitial and telomeric heterochromatin of lake trout, including that of the X chromosome, appears to comprise sequences belonging to the MboI/BglII family.

摘要

湖红点鲑(Salvelinus namaycush)的染色体含有大量异染色质,位于几条染色体的着丝粒和/或端粒处,包括位于X染色体远端的性别特异性区域。为了进一步研究湖红点鲑的重复DNA,用限制性内切酶AluI对一只雌性湖红点鲑的基因组DNA进行了大小分级(<600 bp),片段被克隆到噬菌体M13中。共分离出42个克隆。通过Southern分析估计湖红点鲑基因组中各个插入片段的相对拷贝数。确定12个克隆为高度重复序列,并选择进行进一步研究。这些克隆的插入片段包含与北极红点鲑(Salvelinus alpinus)报道的AluI/RsaI、EcoRI/DraI、DraI/BstEII和MboI/BglII家族相似的序列。通过荧光原位杂交(FISH)确定了湖红点鲑中这些片段中的几个在染色体上的位置。两个相关的AluI/RsaI序列(A型,约140 bp,B型,约120 bp)显示出不同的杂交情况。A型与所有中着丝粒染色体以及几条近端着丝粒染色体的着丝粒杂交。B型与大多数近端着丝粒染色体的着丝粒杂交。一个与EcoRI/DraI家族同源的序列与几条近端着丝粒染色体的着丝粒杂交。与DraI/BstEII家族部分相似的序列与主要的核糖体DNA位点(核仁组织区,NORs)和几个次要的端粒位点杂交。湖红点鲑的间期和端粒异染色质,包括X染色体的异染色质,似乎包含属于MboI/BglII家族的序列。

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