中国水稻科学2017，Vol.31Issue(5)：475-482,8.DOI:10.16819/j.1001-7216.2017.7036
水稻着丝粒特异组蛋白CENH3抗体的制备与应用
Preparation and Application of Centromere-Specific Protein CENH3 Antibody in Rice
摘要
Abstract
[Objective] The centromeres of eukaryotic chromosomes are essential for cell division and inheritance of genetic information. The centromere is a complex composed of centromeric proteins and DNA motifs. In particular, a centromere-specific histone 3 variant, referred to CENH3, is a key centromere-specific protein for centromeric chromatin. So the preparation of CENH3 antibody is necessary for the research of centromeric structure and function.[Method]We designed a short-peptide for the rabbit immune experiment and prepared CENH3 antibody in rice. ELISA and protein immunofluorescence(IF) assay were performed to detect the effectiveness and availability of CENH3 antibody.[Result] The effective dilution of CENH3 antibody was detected at 1:400000. Through IF analysis, obvious signals were detected in the centromeric region of each chromosome in rice and maize. DNA associated with CENH3 was selectively immunoprecipitated in chromatin immunoprecipitation assay(ChIP). After PCR amplification and FISH(fluorescence in situ hybridization) analysis, the results showed ChIP-DNA was located in functional centromere in rice.[Conclusion] These results demonstrated that CENH3 antibody prepared in this study can meet the requirement of related experiments in the research of centromere, and the antibody was prepared successfully.关键词
水稻/着丝粒特异组蛋白/染色质免疫沉淀/蛋白免疫荧光Key words
rice/centromere-specific histone/chromatin immune-precipitation assay/immunofluorescence分类
生物科学引用本文复制引用
薛超,张融,郭瑞,刘帅,刘晓宇,沈明晨,邓世峰,龚志云..水稻着丝粒特异组蛋白CENH3抗体的制备与应用[J].中国水稻科学,2017,31(5):475-482,8.基金项目
国家自然科学基金面上项目(3137021) （3137021）
江苏省"青蓝工程"项目. （）
