ORGANIZATION OF SEA-URCHIN HISTONE GENES
ORGANIZATION OF SEA-URCHIN HISTONE GENES
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DOI：
10.1016/0092-8674(75)90185-3
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发表时间：
1975-01-01
期刊：
影响因子：
64.5
通讯作者：
COHEN, SN
中科院分区：
文献类型：
--
作者：
KEDES, LH;COHN, RH;COHEN, SN
用EcoRI和Hind III限制性内切核酸酶切割的总海胆DNA的蔗糖梯度分析以及通过与组蛋白mRNA杂交鉴定组蛋白编码基因序列已经阐明了组蛋白基因重复单元的基本组织。这些数据，再加上纯化的核酸内切酶切割的海胆组蛋白 DNA 的电泳分析以及与从大肠杆菌中克隆的构建的质粒嵌合体的真核片段转录的 cRNA 杂交获得的结果，表明编码单个组蛋白的几个 DNA 序列混合在一个 7 kb 的重复单元中。用EcoRI切割总海胆DNA产生2.2和4.8kb片段，其与两个克隆片段同源，并且包含在7kb Hind III片段中。用两种酶的切割表明2.2kb EcoRI片段含有距离末端0.15-0.2kb的Hind III位点。嵌合质粒DNA和从海胆胚胎多核糖体分离的纯化的个体mRNA之间的RNA/DNA杂交已被用于将编码序列分配给组蛋白DNA重复单元的2.2或4.8kb区域。提出了组蛋白基因图谱。
Sucrose gradient analysis of total sea urchin DNA cleaved with the EcoRl and Hind Ill restriction endonucleases and identification of histone coding gene sequences by hybridization with histone mRNA have elucidated the basic organization of the histone gene repeat unit. These data, plus results obtained by electrophoretic analysis of purified endonuclease-cleaved sea urchin histone DNA and hybridization with cRNA transcribed from the eucaryotic segment of constructed plasmid chimeras cloned in E. coli, show that the several DNA sequences coding for individual histone proteins are intermingled in a 7 kilobase (kb) repeat unit. Cleavage of total sea urchin DNA with EcoRl produces 2.2 and 4.8 kb fragments which are homologous with the two cloned fragments, and which are contained in a 7 kb Hind III fragment. Cleavage with both enzymes reveals that the 2.2 kb EcoRl fragment contains a Hind III site 0.15-0.2 kb from an end. RNA l DNA hybridization between chimeric plasmid DNA and purified individual mRNAs isolated from sea urchin embryo polyribosomes has been used to assign coding sequences to either the 2.2 or 4.8 kb region of the histone DNA repeat unit. A map of the histone genes is proposed.