摘要
为了研究猪圆环病毒4型(porcine circovirus type 4,PCV4)Cap蛋白功能,本研究构建了其原核表达质粒pCzn1-Cap,并对其进行了诱导表达和蛋白纯化。以纯化的重组蛋白His-Cap作为免疫原免疫BALB/c小鼠,并将SP2/0骨髓瘤细胞与免疫后的小鼠脾细胞融合,通过ELISA筛选得到了1株能够稳定、可持续性分泌与重组蛋白His-Cap有较高特异性反应的单克隆抗体的杂交瘤细胞株,命名为3E6G3。经测定,单克隆抗体3E6G3的抗血清效价可达1∶512000。PCV4 Cap蛋白单克隆抗体的制备将为该蛋白功能的研究和PCV4感染的快速诊断及防控奠定基础。
In order to study the function of the Cap protein of porcine circovirus type 4(PCV4),the prokaryotic expression plasmid pCzn1-Cap was constructed,and the recombinant proteins were induced and purified.BALB/c mice were immunized with purified recombinant protein His-Cap as immunogens,and the immunized mice spleen cells were fused with SP2/0 myeloma cells.Finally,a stable and sustainable monoclonal antibody cell line 3E6G3 with a high specific response to recombinant protein His-Cap was obtained by ELISA screening.The antiserum titer of monoclonal antibody 3E6G3 could reach 1∶512000.The preparation of the monoclonal antibody against PCV4 Cap protein will lay a foundation for the study of the function of the protein and the rapid diagnosis and control of PCV4 infection.
作者
阮青婷
张琦
龚紫凤
农可懿
覃婷
梁小妹
吕其壮
RUAN Qing-ting;ZHANG Qi;GONG Zi-feng;NONG Ke-yi;QIN Ting;LIANG Xiao-mei;LüQi-zhuang(College of Biology&Pharmacy,Yulin Normal University,Yulin 537000,China;Guangxi Key Laboratory of Agricultural Resources Chemistry and Biotechnology,Yulin 537000,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2021年第12期1554-1560,共7页
Chinese Veterinary Science
基金
国家自然科学基金项目(31860708)
广西自然科学基金项目(2017GXNSFBA198025)
玉林师范学院高层次人才科研启动基金项目(G2021ZK12)
玉林师范学院大学生创新创业训练计划立项项目(202110606040)。
关键词
猪圆环病毒4型
CAP蛋白
原核表达
单克隆抗体
porcine circovirus type 4(PCV4)
Cap protein
prokaryotic expression
monoclonal antibody
