奶牛妊娠相关糖蛋白BoPAG16的表达及其结构功能预测  

Eukaryotic expression,structure and function prediction of pregnancy-associated glycoprotein BoPAG16 in dairy cows

作　　者：刘磊[1] 陈柏桥 柳玉 李瑞 陈珊珊 尹茉莉 王会岩[1] LIU Lei;CHEN Bai-qiao;LIU Yu;LI Rui;CHEN Shan-shan;YIN Mo-li;WANG Hui-yan(Jilin Collaborative Innovation Center for Antibody Engineering,Jilin Medical University,Jilin 132013,China;College of Medicine Laboratory,Jilin Medical University,Jilin 132013,China)

机构地区：[1]吉林医药学院,吉林省抗体工程科技协同创新中心,吉林吉林132013 [2]吉林医药学院检验学院,吉林吉林132013

出　　处：《生物技术》2022年第2期135-140,170,共7页Biotechnology

基　　金：吉林省科技发展计划项目(20180623045TC,20190301088NY);吉林市科技创新发展计划项目(201830853);吉林省大学生创新创业训练计划项目(202013706053)。

摘　　要：[目的]为了获得奶牛妊娠相关糖蛋白16(bovin pregnancy associated glycoproteins 16,BoPAG16),并对该蛋白进行结构和功能分析。[方法]从荷斯坦奶牛胎盘子叶中扩增BoPAG16基因,并用构建真核表达载体pcDNA3.1-Flag-BoPAG16,转染至293F细胞中,用SDS-PAGE和Western Blotting检测表达效果,用Flag-tag亲和层析柱纯化蛋白,并对BoPAG16基因编码的蛋白进行结构和功能预测分析。[结果]BoPAG16基因在293F细胞中成功表达,大小为56 kDa,纯化后纯度达到92%。预测BoPAG16蛋白N端15个氨基酸为信号肽,6个糖基化位点和13个B细胞抗原表位。[结论]成功表达并纯化奶牛妊娠相关糖蛋白BoPAG16,BoPAG16蛋白有6个糖基化位点和13个B细胞抗原表位。[Objective] In order to obtain BoPAG16(bovine Pregnancy associated glycoprotein 16),and we carried out predict and analyze analysis.[Method] The total RNA extracted from placenta cotyledons of Holstein cows and connected with pcDNA3.1 vector by T4 DNA ligase.The expression vector pcDNA3.1-boPAG16 was transfected into 293 F cells.SDS-PAGE and Western Blotting were performed to detect the expression of the BoPAG16.The expression product was purified by Flag-tag and carried out predict,analyze analysis of BoPAG16 protein.[Result]The boPAG16 gene was successfully expressed in 293 F cell,the protein size is 56 kDa.The BoPAG16 reached above 92% after purification by Flag-tag column chromatography.The BoPAG16 protein had a signal peptide composed of 15 amino acids,six glycosylation sites and thirteen B cell epitopes.[Conclusion] BoPAG16 protein was successfully expressed and purified which six glycosylation sites and thirteen B cell epitopes.

关 键 词：牛妊娠相关糖蛋白 奶牛 胎盘子叶 真核表达 纯化 

分 类 号：Q786[生物学—分子生物学]