中国免疫学杂志2024，Vol.40Issue(1)：72-77,6.DOI:10.3969/j.issn.1000-484X.2024.01.009
多态性BoLA Ⅰα1α2与恒定链结合及在真核细胞共定位
Polymorphic BolA Ⅰα1α2 binds to constant chains and co-locates in eukaryotic cells
摘要
Abstract
Objective:To demonstrate the polymorphism of α chain of bovine major histocompatibility complex(BoLA)classⅠmolecule and domain binding constant chain(Ii).Methods:Total 75 BoLA Iα genes were obtained from three Huaibei cattle and analyzed by molecular biology software;the genes of typical BoLA Iα domains and Ii were cloned,and then inserted into prokaryotic expression plasmid.After induced protein expression;the domains of BoLA Ⅰα chain binding to Ii were detected by pull-down meth-od and Western blot.The recombinant eukaryotic expression plasmids were constructed and the co-localization of BoLA Iα segments with Ii was observed by laser confocal microscopy.Results:Firstly,it was found that there were at least 5 kinds of BoLA Iα in the cloned gene sequence,which were highly polymorphic and they were mainly distributed in the antigen peptide binding region(PBR)of BoLA Ⅰ(α1α2)and cytoplasmic region.Secondly,the prokaryotic recombinant plasmids containing BoLA Ⅰα1α2α3,BoLA Ⅰα1α2 or BoLA Ⅰα 3 were constructed,then they were respectively induced to express and purified,in which,the BoLA Ⅰα1α2α3 and BoLA Ⅰα1α2 had the activity of binding to Ii.Finally,in 293T cells BoLA Ⅰα1α 2α3 or BoLA Ⅰα1α2 was found that could co-localize with Ii,while a single BoLA Ⅰα3 could not.Conclusion:BoLA Ⅰα gene is highly polymorphic.BoLA Ⅰα1α2 is a func-tional fragment that binds to Ii and co-locates intracellular.关键词
BoLA/高度多态性/Ii/结合/共定位Key words
BoLA/Highly polymorphic/Ii/Combination/Co-localization分类
农业科技引用本文复制引用
陈芳芳,于凤梅,刘翠艳,桂亚萍,李锦春..多态性BoLA Ⅰα1α2与恒定链结合及在真核细胞共定位[J].中国免疫学杂志,2024,40(1):72-77,6.基金项目
国家自然科学基金项目(31572496) （31572496）
安徽省自然科学基金项目(2108085MC116) （2108085MC116）
安徽省教育厅重点项目(KJ2020A0124). （KJ2020A0124）
