中国兽医杂志2023，Vol.59Issue(12)：49-53,5.
赤羽病病毒Gcaa465～704的杆状病毒表达和单克隆抗体制备
Baculovirus Expression and Monoclonal Antibody Preparation of Akabane Virus Gcaa465-704
摘要
Abstract
To prepare monoclonal antibody(mAb)against the glycoprotein Gc of Akabane virus(AKAV),the gene sequence of AKAV Ge protein from amino acid 465 to 704 was codon-optimized and synthesized into the pFastBac HTB vector.After blue-white screening,recombinant AKAV Gcaa465-704 was produced using the baculovirus expression system in insect cells.Then,Gcaa465-704 was purified and used for the immunization of mice.Positive hybridoma cells secreting mAb were obtained through cell fusion and subcloning,and the mAb characteristics were identified by Western blot and indirect immunofluorescence assay(IFA).The results showed that the hybridoma cell line 4D1 stably secreting mAb against AKAV Gcaa465-704 was successfully screened.Western blot results demonstrated that the mAb 4D1 could specifically recognize AKAV Gcaa465-704.And IFA results showed that mAb 4D1 could react with AKAV-infected BHK21 cells.This study successfully prepared mAb against AKAV Gcaa465-704,which provides technical support for further research on the biological functions of AKAV Gc and the establishment of AKAV detection methods.关键词
赤羽病病毒/Gc蛋白/杆状病毒表达/单克隆抗体Key words
Akabane virus/Gc protein/baculovirus expression/monoclonal antibody分类
农业科技引用本文复制引用
魏方,陈冬杰,邓俊花,王晶晶,吴绍强..赤羽病病毒Gcaa465～704的杆状病毒表达和单克隆抗体制备[J].中国兽医杂志,2023,59(12):49-53,5.基金项目
基本科研业务费项目(2022JK11) （2022JK11）
"十四五"国家重点研发计划(2022YFD1800505) （2022YFD1800505）
