动物医学进展2026，Vol.47Issue(5)：35-42,8.
牛源副结核分枝杆菌MAP 0862蛋白的原核表达及抗体制备
Prokaryotic Expression of Mycobacterium avium subsp.paratuberculosis Protein MAP 0862 of Cow Origin and Antibody Preparation
摘要
Abstract
In order to prepare the monoclonal antibody specific to Mycobacterium avium subsp.paratubercu-losis(MAP)protein MAP 0862 and the polyclonal antibody against MAP,MAP 0862 expression vector pET-28a(+)-0862 was constructed,and the recombinant protein was induced to be expressed by IPTG,and the recombinant protein MAP 0862 was successfully expressed as identified by SDS-PAGE,and the mice were immunized with it after purification,and then immunized against BALB/c mice.MAP 0862 mouse-derived monoclonal antibody was prepared by hybridoma technique,and the Western blot results showed that MAP 0862 mouse-derived monoclonal antibody could specifically bind to MAP 0862,and the antibody titer was 1∶12 800 by indirect ELISA method;Formaldehyde inactivated MAP strain immunized New Zealand white rabbits to prepare MAP rabbit polyclonal antibody,and the Western blot results showed that MAP rabbit-derived polyclonal antibody could bind to MAP 0862,and the Western blot results showed that MAP rabbit-derived polyclonal antibody could bind to MAP 0862 specifically.Western blot results showed that the MAP rabbit polyclonal antibody could bind specifically to MAP,and the potency of the antibody was 1∶3 200 by indirect ELISA.In conclusion,the mouse monoclonal antibody to MAP 0862 and the rabbit polyclonal antibody to MAP prepared in the present study have good reactivity and specificity,and provide a theoretical basis for the study of the biological characteristics of MAP 0862,the establishment of a method of detecting MAP and the development of a vaccine.关键词
副结核分枝杆菌/MAP 0862/原核表达/单克隆抗体/多克隆抗体Key words
Mycobacterium avium subsp.Paratuberculosis/MAP 0862/prokaryotic expression/Monoclonal antibody/Polyclonal antibody分类
农业科技引用本文复制引用
杨文洁,谢梦圆,张勇,付婧,丁宁,白海鑫,郝彦儒,徐晓静..牛源副结核分枝杆菌MAP 0862蛋白的原核表达及抗体制备[J].动物医学进展,2026,47(5):35-42,8.基金项目
内蒙古自治区兴安盟科技合作项目(MBHZ2025001) （MBHZ2025001）
内蒙古自治区呼和浩特市科技创新领域人才项目(2022RC-联合体-1) （2022RC-联合体-1）
内蒙古自治区科技计划项目(2023YFDZ0058) （2023YFDZ0058）
内蒙古自治区科技重大专项(2020ZD0006) （2020ZD0006）
