摘要
【目的】在昆虫细胞表达猪圆环病毒2型(PCV2)衣壳蛋白(Cap),并对表达的重组蛋白~*Cap进行免疫原性分析。【方法】将含有PCV2-ORF2基因的重组杆状病毒质粒Bacmid-ORF2转染昆虫细胞进行蛋白表达试验,再用纯化的~*Cap和质粒pcDNA3.1-ORF2分别免疫小鼠,比较二者的免疫效果。【结果】由PCV2-ORF2基因编码的Cap结构蛋白在昆虫细胞中正确表达,相对分子质量约为32 000。~*Cap能够被PCV2阳性血清所识别,具有良好的抗原性。~*Cap免疫组在细胞免疫水平及体液免疫水平的免疫效果均优于pcDNA3.1-ORF2免疫组。【结论】重组蛋白~*Cap可替代全病毒,具有PCV2疫苗潜在的应用价值。研究结果为后续的PCV2病毒分子诊断、亚单位疫苗以及分子生物学等研究奠定试验基础。
【Objective】To express the capsid protein（Cap） of porcine circovirus type 2（PCV2） in insect cells and analyze the immunogenicity of recombinant protein.【Method】The established recombinant bacmid containing fused ORF2 gene（Bacmid-ORF2） was transfected into insect cells for protein expression test, and then the purified ＊Cap and pcDNA3.1-ORF2 plasmid were immunized mice respectively to compare their immunological effects.【Result】The Cap structural protein encoded by PCV2-ORF2 gene was correctly expressed in insect cells, and the relative molecular mass was about 32 000. ＊Cap could be recognized by PCV2 positive serum and had good antigenicity. The immue effect of ＊Cap group was better than that of pcDNA3.1-ORF2 group in both cellular immunity and humoral immunity levels.【Conclusion】The recombinant protein＊Cap can replace the whole virus and may have potential application values of PCV2 vaccinum. These resultslay the foundation for the study of molecular diagnosis, subunit vaccine development and molecular biology research of PCV2.
作者
徐铮
马晓莉
林彦星
董建国
王磊
刘燕玲
刘清神
宋长绪
XU Zheng1, MA Xiaoli2, LIN Yanxing3, DONG Jianguo1, WANG Lei1, LIU Yanling1, LIU Qingshen1, SONG Changxu1(1. National Engineering Research Center for Breeding Swine Industry/College of Animal Science, South China Agricultural University, Guangzhou 510642, China; 2. Laboratory Animal Center, South China Agricultural University, Guangzhou 510642, China; 3. Inspection and Quarantine Center for Animals and Plants, Shenzhen Entry-exit Inspection and Quarantine Bureau, Shenzhen 518045, Chin)
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2018年第3期1-5,共5页
Journal of South China Agricultural University
基金
"十二五"农村领域国家科技计划(2015BAD12B02-5)
现代农业产业共性技术创新团队项目(2016LM2150)
广东省科技计划(2016A020212021
2015B020230004)
广州市科技计划(201508020062)
作者简介
徐铮（1979-），男，助理研究员，硕士，E—mail：stonezen@scau.edu.cn；;通信作者：宋长绪（1965-），男，研究员，博士，E-mail：xsong@scau.edu.cn
