{"title":"重组组蛋白纯化及单核小体组装","authors":"Yiming Zhao, Jingjun Hong","doi":"10.1002/cpz1.70155","DOIUrl":null,"url":null,"abstract":"<p>The nucleosome is the basic unit of chromatin. Each nucleosome is composed of a little less than two turns of DNA wrapped around a set of eight proteins called histones, which are known as a <span>histone octamer</span>. Each histone octamer is composed of two copies each of the histone proteins <span>H2A</span>, <span>H2B</span>, <span>H3</span>, and <span>H4</span>. Nucleosomes play an important role in gene expression and regulation. While previous protocols use HPLC (high-performance liquid chromatography) to purify each histone and, after octamer reconstitution, subsequent FPLC (fast protein liquid chromatography) to purify the octamers. Here we present a method to carry out octamer reconstitution and mononucleosome assembly with FPLC only. This basic protocol describes a procedure for histone purification from <i>Escherichia coli (E. coli)</i>, their subsequent reconstitution as octamers, and assembly into mononucleosomes <i>in vitro</i>. Through this protocol, histone octamers and mononucleosomes can be reconstituted on a large scale without the use of HPLC. Overall, this protocol describes an effective new method for the reconstitution of octamers and assembly of mononucleosomes. © 2025 Wiley Periodicals LLC.</p><p><b>Basic Protocol</b>: Purification of recombinant histones and mononucleosome assembly</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 5","pages":""},"PeriodicalIF":2.2000,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Purification of Recombinant Histones and Mononucleosome Assembly\",\"authors\":\"Yiming Zhao, Jingjun Hong\",\"doi\":\"10.1002/cpz1.70155\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>The nucleosome is the basic unit of chromatin. Each nucleosome is composed of a little less than two turns of DNA wrapped around a set of eight proteins called histones, which are known as a <span>histone octamer</span>. Each histone octamer is composed of two copies each of the histone proteins <span>H2A</span>, <span>H2B</span>, <span>H3</span>, and <span>H4</span>. Nucleosomes play an important role in gene expression and regulation. While previous protocols use HPLC (high-performance liquid chromatography) to purify each histone and, after octamer reconstitution, subsequent FPLC (fast protein liquid chromatography) to purify the octamers. Here we present a method to carry out octamer reconstitution and mononucleosome assembly with FPLC only. This basic protocol describes a procedure for histone purification from <i>Escherichia coli (E. coli)</i>, their subsequent reconstitution as octamers, and assembly into mononucleosomes <i>in vitro</i>. Through this protocol, histone octamers and mononucleosomes can be reconstituted on a large scale without the use of HPLC. Overall, this protocol describes an effective new method for the reconstitution of octamers and assembly of mononucleosomes. © 2025 Wiley Periodicals LLC.</p><p><b>Basic Protocol</b>: Purification of recombinant histones and mononucleosome assembly</p>\",\"PeriodicalId\":93970,\"journal\":{\"name\":\"Current protocols\",\"volume\":\"5 5\",\"pages\":\"\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2025-05-22\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.70155\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols","FirstCategoryId":"1085","ListUrlMain":"https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.70155","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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