深圳大学学报（理工版）2025，Vol.42Issue(5)：513-520,8.DOI:10.3724/SP.J.1249.2025.05513
利用SPIDER和IP-MS技术鉴定小立碗藓着丝粒关键蛋白
Key centromere proteins in Physcomitrella patens identified via SPIDER and IP-MS
摘要
Abstract
Plant artificial chromosomes play a crucial role in synthetic biology,wherein key centromere proteins represent essential components in synthesizing these chromosomes.However,the identification of centromere proteins in plants remains limited.To identify more centromere proteins,this study utilized Physcomitrella patens-specific centromere histone(PpCENH3)as a bait molecule and employed immunoprecipitation-mass spectrometry(IP-MS)to systematically identify 13 potential centromere-associated proteins.Further analysis of conserved centromere monomer repeats sequences in P.patens led to the design of a 72-bp(base pair)bait deoxyribonucleic acid.Using the latest proximity labeling technology,specific pupylation as IDEntity reporter(SPIDER),we successfully identified 27 centromere-associated proteins.Compared to IP-MS,SPIDER identified 16 additional potential key centromere proteins,demonstrating its superiority over traditional IP-MS technology in identifying key centromere proteins.Combining both methods,we identified a total of 29 centromere-associated proteins,with 11 proteins identified by both techniques.These findings not only provides new insights into the structure and function of plant centromeres but also establishes a foundation for the development of stable artificial plant chromosomes.关键词
植物人工染色体/小立碗藓/邻近标记技术/免疫沉淀-质谱联用/着丝粒关键蛋白/着丝粒单体序列Key words
plant artificial chromosomes/Physcomitrella patens/proximity labeling technology/immunoprecipitation-mass spectrometry(IP-MS)/key centromere protein/centromere monomer sequence分类
农业科技引用本文复制引用
刘学东,翁玉明,王铭亮,莫蓓莘..利用SPIDER和IP-MS技术鉴定小立碗藓着丝粒关键蛋白[J].深圳大学学报（理工版）,2025,42(5):513-520,8.基金项目
National Basic Research Program of China(2019YFA0903902) （2019YFA0903902）
National Natural Science Foundation of China(32402310) （32402310）
Basic and Applied Basic Research Foundation of Guangdong(2023A1515110407) 国家重点基础研究发展计划资助项目(2019YFA0903902) （2023A1515110407）
国家自然科学基金资助项目(32402310) （32402310）
广东省基础与运用基础研究基金项目(2023A1515110407) （2023A1515110407）
