中国医科大学学报2024，Vol.53Issue(6)：495-500,6.DOI:10.12007/j.issn.0258-4646.2024.06.003
β淀粉样蛋白1-42质粒的构建及其与钙调蛋白的结合作用
Construction of Aβ1-42 plasmid and its binding to calmodulin
摘要
Abstract
Objective To investigate the involvement of calmodulin(CaM)in the pathogenesis of Alzheimer disease(AD)and the mechanism by which CaM binds to amyloid-β(Aβ).Methods The hub genes expressed in AD and predicted to be the target proteins for AD prevention and treatment were obtained using bioinformatics methods.The GST-Aβ1-42 recombinant plasmid was constructed through genetic recombination and was then sequenced.The recombinant plasmids were identified using agarose gel electrophoresis,while the extracted and purified GST-Aβ1-42 fusion protein was confirmed using SDS-PAGE gel electrophoresis.GST pull-down assay was used to detect the interaction between GST-Aβ1-42 protein and CaM,expressed in the plasmid.Results The top 20 hub genes in degree ranking were obtained.The DNA sequencing results of the plasmid proved that the recombinant plasmid was successfully constructed.The agarose gel electrophoresis results indicated that the fragment digested by the enzyme was similar to the molecular weight of the Aβ1-42 gene seg-ments,further proving the successful construction of the recombinant plasmid.Binding of GST-Aβ1-42 protein to CaM in a concentration dependent manner was revealed through the GST pull down experiment.Conclusion The GST-Aβ1-42 recombinant plasmid is success-fully constructed and is shown to bind to CaM.关键词
β淀粉样蛋白/质粒构建/钙调蛋白/结合能力/活性鉴定Key words
amyloid-β/plasmid construction/calmodulin/binding ability/activity identification分类
医药卫生引用本文复制引用
齐爽,孙旋旋,王绮萱,贺意婷,李佳睿,苏敬阳,郝丽英..β淀粉样蛋白1-42质粒的构建及其与钙调蛋白的结合作用[J].中国医科大学学报,2024,53(6):495-500,6.基金项目
辽宁省大学生创新训练项目(S202210159031,S20231015944,S202310159018) （S202210159031,S20231015944,S202310159018）
医学电生理学教育部重点实验室开放基金(KeyME-2019-07) （KeyME-2019-07）
