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人类及其他灵长类基因组中长散布重复DNA的KpnI家族。

KpnI families of long, interspersed repetitive DNAs in human and other primate genomes.

作者信息

Shafit-Zagardo B, Maio J J, Brown F L

出版信息

Nucleic Acids Res. 1982 May 25;10(10):3175-93. doi: 10.1093/nar/10.10.3175.

Abstract

KpnI restriction of DNAs from all anthropoid primates studied releases a conspicuous series of segments representing families of long, interspersed repetitive DNAs termed here the KpnI 1.2, 1.5, 1.8 and 1.9 kb families. Human KpnI 1.2 to 1.9 kb segments representative of these families were isolated and separately cloned in the KpnI site of a plasmid pBK5, specially constructed for this purpose. The KpnI clones did not cross-hybridize with cloned, primate alphoid sequences, suggesting that the KpnI families represent sequences separate and distinct from the alphoid DNAs. Secondary restriction analyses of cloned KpnI segments demonstrated microheterogeneity among individual members within the same KpnI family. Autoradiograms of capuchin monkey, AGM and human DNA cleaved with HaeIII, AluI or RsaI and hybridized to various cloned human KpnI sequences demonstrated a remarkable conservatism and relative simplicity in the organization of the KpnI families in the genomes of these widely divergent primates. The KpnI 1.2 kb and 1.5 kb families occur in high frequency (15%) among all plaques in two recombinant human genome libraries. Evidence is presented suggesting that the bulk of the KpnI families occur in the genome as clusters or congeries of higher molecular weight segments (greater than 2 kb) containing sequences homologous to the low molecular weight segments (1.2 to 1.9 kb).

摘要

对所有被研究的灵长类动物的DNA进行KpnI酶切,会释放出一系列显著的片段,这些片段代表了长散在重复DNA家族,在这里被称为KpnI 1.2、1.5、1.8和1.9 kb家族。分离出代表这些家族的人类KpnI 1.2至1.9 kb片段,并分别克隆到专门为此构建的质粒pBK5的KpnI位点中。KpnI克隆与克隆的灵长类α卫星序列不发生交叉杂交,这表明KpnI家族代表的序列与α卫星DNA是分开且不同的。对克隆的KpnI片段进行的二次酶切分析表明,同一KpnI家族的个体成员之间存在微异质性。用HaeIII、AluI或RsaI切割的卷尾猴、非洲绿猴和人类DNA的放射自显影片,与各种克隆的人类KpnI序列杂交,结果表明,在这些亲缘关系甚远的灵长类动物的基因组中,KpnI家族的组织具有显著的保守性和相对简单性。在两个重组人类基因组文库的所有噬菌斑中,KpnI 1.2 kb和1.5 kb家族以高频率(15%)出现。有证据表明,KpnI家族的大部分在基因组中以高分子量片段(大于2 kb)的簇或集合形式存在,这些高分子量片段包含与低分子量片段(1.2至1.9 kb)同源的序列。

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