畜牧兽医学报2012，Vol.43Issue(8)：1266-1272,7.
猪繁殖与呼吸综合征病毒GP3/GP5/M真核表达载体的构建及其体液免疫应答
Construction and Humoral Immune Response of Recombinant Plasmid Co-Expressing GP3, GP5 and M Proteins of Porcine Reproductive and Respiratory Syndrome Virus
王凡 1刘建斌 2祝秀梅 1吕志慧 1马全英 1杜平 1刘学荣 1牟克斌1
作者信息
- 1. 中农威特生物科技股份有限公司,兰州730046
- 2. 中国农业科学院兰州畜牧与兽药研究所,兰州730050
- 折叠
摘要
Abstract
The aim of this study was to construct a recombinant eukaryotic plasmid co-expressing GP3, GP5 and M proteins of porcine reproductive and respiratory syndrome virus (PRRSV). GP3, GP5 and M protein gene fragments of PRRSV LN strain were amplified by PCR and cloned into vector pcDNA3. 1 (+) to produce the recombinant plasmid pcDNA3. 1-GP3-GP5-M. The plasmid was transfected into COS7 cells, and immunological responses to the plasmid were investigated in BALB/c mice and piglets. The recombinant plasmid pcDNA3. 1-GP3-GP5-M was transfected into COS7 cells, PCR identification and indirect immunofluorescence assay proved that GPS, GP5-M gene of PRRSV were expressed in COS7 cells. The result of Western blotting , showed that the GP3 and GP5 and M proteins were co-expressed and formed fusion protein. The BALB/c mice were injected with recombinant plasmid pcDNA3. 1-GP3-GP5-M to evaluate the induced immunological responses in vivo. The specific detectable anti-PRRSV neutralizing antibod-ies were produced in the vaccinated mice at 2 weeks and reached a peak 1:32 at 8 weeks after primary vaccination. In addition, it was observed that the 1:4-1:8 neutralizing antibodies of the vaccinated piglets. The results showed that the recombinant plasmid pcDNA3. 1-GP3-GP5-M has been constructed successfully, and the recombinant plasmid has well immunity, the recombinant plasmid could be a basis to eukaryotic vector vaccine for PRRSV infection.
关键词
猪繁殖与呼吸综合征病毒(PRRSV)/GP3蛋白/GP5蛋白/M蛋白Key words
porcine reproductive and respiratory syndrome virus (PRRSV)/ GP3 protein/ GP5 protein/ M protein分类
农业科技引用本文复制引用
王凡,刘建斌,祝秀梅,吕志慧,马全英,杜平,刘学荣,牟克斌..猪繁殖与呼吸综合征病毒GP3/GP5/M真核表达载体的构建及其体液免疫应答[J].畜牧兽医学报,2012,43(8):1266-1272,7.
