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通过酵母中的基因筛选对盘基网柄菌编码核小体核心组蛋白H3的cDNA进行分子克隆。

Molecular cloning of a cDNA encoding the nucleosome core histone H3 from Dictyostelium discoideum by genetic screening in yeast.

作者信息

Bukenberger M, Horn J, Dingermann T, Dottin R P, Winckler T

机构信息

Hunter College, Department of Biological Sciences, New York, NY 10021, USA.

出版信息

Biochim Biophys Acta. 1997 May 2;1352(1):85-90. doi: 10.1016/s0167-4781(97)00029-8.

Abstract

The one-hybrid method for genetic screening in yeast was used to search a Dictyostelium discoideum cDNA library for DNA-binding proteins that interact with the C-module of the Dictyostelium repetitive element. The C-module was formerly shown to contain two high affinity, sequence-specific binding sites for a nuclear protein factor of unknown function (CMBF). The bait DNA sequence was bound in vivo by a cDNA-encoded protein whose derived amino acid sequence showed high homology to nucleosome core histone H3, but not to partially available CMBF sequences. The D. discoideum histone H3 homolog is encoded by a single gene and shows significant sequence variation at the amino terminus of the protein, including a triple-serine insertion not found in any other histone H3.

摘要

利用酵母中的单杂交方法对盘基网柄菌cDNA文库进行筛选,以寻找与盘基网柄菌重复元件C模块相互作用的DNA结合蛋白。先前已表明C模块含有两个高亲和力、序列特异性的结合位点,可与功能未知的核蛋白因子(CMBF)结合。诱饵DNA序列在体内与一种由cDNA编码的蛋白结合,该蛋白的推导氨基酸序列与核小体核心组蛋白H3具有高度同源性,但与部分已知的CMBF序列无同源性。盘基网柄菌组蛋白H3同源物由单个基因编码,在蛋白的氨基末端表现出显著的序列变异,包括在任何其他组蛋白H3中均未发现的三联丝氨酸插入。

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