摘要
仔猪轮状病毒腹泻严重危害养猪业,为了开发亚单位疫苗,本课题组前期构建了猪轮状病毒SB-1A株截短的VP8^*蛋白(△VP8^*,64~223位氨基酸)原核表达载体pET28a—SB1A△VP8^*。为了进一步提高亚单位疫苗的免疫原性,将破伤风毒素的通用T细胞表位P2引入重组亚单位蛋白,构建重组载体pET28a—P2-SB-IA△VP8^*。P2-S13-1A△VP8^*和S13-1A△VP8^*在大肠杆菌中表达并纯化。重组蛋白与氢氧化铝佐剂混合后经肌肉注射免疫Balb/c小鼠,利用间接ELISA法检测免疫小鼠的血清抗体水平。结果显示:P2-S13-1A△VP8^*和S昏1A△VP8^*在大肠杆菌中得到了高水平、可溶性表达;重组蛋白P2-SB-1A△VP8^*诱导的血清抗体水平显著高于SB-1A△VP8^*,说明P2可以增强SB-1A△VP8^*的免疫原性,为日后开发猪轮状病毒亚单位疫苗奠定了基础。
Porcine rotavirus is a major cause of piglet diarrhea,resulting in serious burden to animal husbandry. In our previous research,the truncated VP8* protein (△VP8* ,64-223 aa ) prokaryotic expression vector pET28a-SB-1A/△VP8* of porcine rotavirus SB-1A strain was constructed. In an attempt to improve the immunogenicity of subunit vaccine protein, universal tetanus toxin T-cell epitope P2 was introduced to the upstream of truncated gene, constructed recombinant vector pET28a-P2-SB-1A △VP8* ,followed by the induction of P2-SBqA △VP8* recombinant protein in Escherichia coli and purification of recombinant protein P2-SB-1A △VP8* , as well as SB-1A △VP8*. Balb/c mice were immunized intramuscularly with recombinant proteins formulated with aluminum hydroxide. The levels of sera IgG was measured by indirect enzyme-linked immunosorbent assay (ELISA) upon the optimization. The results showed that the two recombinant proteins were soluble and high yield. The serum antibody levels induced by recombinant protein P2- SB-1A △VP8* were extremely significantly higher than that by SB-1A △VP8,indicating that P2 can enhance the immunogenicity of SB-1A △VP8* and P2-SB-1A △VP8* recombinant protein will be potential of the porcine rotavlrus subunit vaccine candidate.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2016年第2期191-195,共5页
Chinese Journal of Veterinary Science
基金
国家自然科学青年基金资助项目(31201909)
黑龙江博士后科研启动资助项目(LBH-Q13134)
黑龙江省自然科学青年基金资助项目(QC2013C028)