阿拉丁SCI论文搜索工具

Current Protocols · 2025 ·Vol. 5 ·期 5 ·e70151 · IF 2.2

Co-Expression and Purification of the Complex of Single-Chain H2B-H2A.Z and Histone Chaperone YL1

Yue Liu,Wansen Tan,Jingjun Hong

Chromatin remodeling serves as a critical mechanism for gene regulation, enabling the replacement of canonical histones with their variants through the assistance of histone chaperones. Our study focused on the process by which the histone variant H2A.Z replaces H2A, mediated by the mammalian SRCAP chromatin remodeling complex subunit hYL1. Previous approaches typically involved separately expressing the two proteins and validating their interaction in vitro . In contrast, we designed an artificial single-chain hH2B-hH2A.Z (hTBZ) construct, co-transformed the hYL1 gene with hTBZ, and co-expressed the proteins in E. coli BL21(DE3). We then performed purification, fast protein liquid chromatography (FPLC), and SDS-PAGE analysis. This approach successfully yielded the complexed proteins, confirming the in vivo interaction between hYL1 and hTBZ. © 2025 Wiley Periodicals LLC. Basic Protocol : Co-transformation of hTBZ/YL1 into BL21 (DE3) competent cells, followed by induction and acquisition of protein

CAS / MDL: 110-18-9 329-98-6 367-93-1 60-00-4 637-50-3 7727-54-0

研究领域: 生物化学 · 分子生物学 · 遗传学

✓ 已验证产品关联 DOI PubMed 高影响因子

查看原文查看 PubMed

Current Protocols · 2025 ·Vol. 5 ·期 7 ·e70179 · IF 2.2

In vitro Nucleosome Assembly Facilitated by the Histone Chaperone SRCAP

Bingyan Yuan,Jingjun Hong

In eukaryotic cells, nucleosomes are octameric structures composed of DNA and histones, and their tandem arrangement ultimately forms chromosomes. Nucleosome assembly is governed in vivo by a series of molecular mechanisms, particularly histone chaperones such as Swr1, which is indispensable for H2A.Z-nucleosome formation. The assessment of nucleosome assembly using micrococcal nuclease (MNase) digestion has been widely accepted. We describe a simple and reproducible protocol for analyzing the chaperone activity of SRCAP (Snf2-related CREBBP activator protein), the histone H2A.Z chaperone. This approach involves purifying a complex of H2A.Z, H2B, and SRCAP via fast protein liquid chromatography (FPLC) only, followed by its assembly with DNA-(H3.1-H4 A15C ) 2 . The formation of nucleosomes is then determined in vitro using electrophoretic mobility shift assay (EMSA) and MNase digestion. This approach can also be adapted to evaluate whether other histone chaperones or regulatory factors promote nucleosome assembly activity. © 2025 Wiley Periodicals LLC. Basic Protocol : In vitro nucleosome assembly facilitated by the histone chaperone SRCAP

研究领域: 分子生物学 · 生物化学 · 细胞生物学 · 表观遗传学

✓ 已验证 DOI PubMed 高影响因子

查看原文查看 PubMed

查找使用阿拉丁产品的科研文献

与阿拉丁产品相关的已发表科研成果

热门产品

Co-Expression and Purification of the Complex of Single-Chain H2B-H2A.Z and Histone Chaperone YL1

In vitro Nucleosome Assembly Facilitated by the Histone Chaperone SRCAP