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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >MUTAGENESIS OF THE HUMAN APOLIPOPROTEIN B GENE IN A YEAST ARTIFICIAL CHROMOSOME REVEALS THE SITE OF ATTACHMENT FOR APOLIPOPROTEIN(A)
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MUTAGENESIS OF THE HUMAN APOLIPOPROTEIN B GENE IN A YEAST ARTIFICIAL CHROMOSOME REVEALS THE SITE OF ATTACHMENT FOR APOLIPOPROTEIN(A)

机译：酵母人工染色体中人类载脂蛋白B基因的突变揭示了载脂蛋白（A）的附着位点

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摘要

Lipoprotein(a) [Lp(a)] is a lipoprotein formed by the disulfide linkage of apolipoprotein (ape) B100 of a low density lipoprotein particle to apolipoprotein(a). Prior studies have suggested that one of the C-terminal Cys residues of apo-B100 is involved in the disulfide linkage of apo-B100 to apo(a). To identify the apo-B100 Cys residue involved in the formation of Lp(a), we constructed a yeast artificial chromosome (YAC) spanning the human apo-B gene and used gene-targeting techniques to change Cys-4326 to Gly. The mutated YAC DNA was used to generate transgenic mice expressing the mutant human apo-B100 (Cys4326Gly). Unlike the wild-type human apo-B100, the mutant human apo-B100 completely lacked the ability to bind to apo(a) and form Lp(a). This study demonstrates that apo-B100 Cys-4326 is required for the assembly of Lp(a) and shows that gene targeting in YACs, followed by the generation of transgenic mice, is a useful approach for analyzing the structure of large proteins coded for by large genes. [References: 30]
机译：脂蛋白（a）[Lp（a）]是通过低密度脂蛋白颗粒的载脂蛋白（ape）B100与载脂蛋白（a）的二硫键形成的脂蛋白。先前的研究表明，apo-B100的C端Cys残基之一与apo-B100与apo（a）的二硫键有关。为了鉴定参与Lp（a）形成的apo-B100 Cys残基，我们构建了跨越人类apo-B基因的酵母人工染色体（YAC），并使用基因靶向技术将Cys-4326更改为Gly。突变的YAC DNA用于生成表达突变型人载脂蛋白B100（Cys4326Gly）的转基因小鼠。与野生型人apo-B100不同，突变型人apo-B100完全缺乏与apo（a）结合并形成Lp（a）的能力。这项研究表明，Apo-B100 Cys-4326是Lp（a）组装所必需的，并且表明，在YAC中靶向基因，然后生成转基因小鼠，是分析大肠埃希菌编码的大蛋白结构的有用方法。大基因。 [参考：30]

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