摘要
为增强表位疫苗的免疫原性,本研究将O型口蹄疫病毒(FMDV)VP1中编码T细胞表位肽(aa 21～aa 40)序列连接于猪细小病毒(PPV)VP2的5'端,并插入到杆状病毒供体质粒pFastBac HTA中,构建成重组转座载体pFastBac-FMDV-VP1-PPV-VP2,转化含有穿梭载体Bacmid的E.coli DH10Bac中,经蓝白菌落筛选获得重组杆状病毒表达质粒rBac-FMDV-VP1-PPV-VP2,转染昆虫草地夜蛾(Sf9)细胞,并在Sf9中进行了表达。电镜观察显示,表达的重组蛋白能够自我组装成病毒样颗粒rPPV∶VLP(FMDV)。用间接免疫荧光试验(IFA)、western blot分析表明,表达产物具有与PPV VP2天然蛋白相似的免疫原性。小鼠免疫实验证实,在低浓度FMDV抗原下能够产生特异性T淋巴细胞增殖反应;而且该病毒样颗粒能诱导机体产生抗FMDV特异性细胞免疫反应。
To improve epitope-vaccine immunogenicity of Food-and-mouth disease virus（FMDV）,The recombinant plasmid pFastBac-FMDV-VP1-PPV-VP2 was constructed by inserting the fusion gene of Porcine parvovirus（PPV） VP2 with the VP1 sequence encoding T-cell epitope（aa21 to aa40） of FMDV type O and transformed into competent E.coli DH10Bac to generate recombinant rBac-FMDV-VP1-PPV-VP2,which was then transfected into insect cell Sf9 with Lipofectamine.Expression product was detected by IFA and western blot.The result showed that the recombinant protein had the similar biological activity to PPV VP2 protein and the self-assembly virus-like particles of rPPV∶VLP（FMDV） could be observed by electron microscopy.The rPPV∶ VLP（FMDV） could induce proliferation of spleen T cells in vaccinated BALB/c and activate FMDV specific cellular immune response.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2010年第11期844-848,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
江苏省农业科技自主创新资金项目(0610808-2)
关键词
猪细小病毒
病毒样颗粒
O型口蹄疫病毒
T细胞表位
porcine parvovirus
virus-like particles
food-and-mouth disease virus type O
T-cell epitope
