表达PRRSV M蛋白重组腺病毒的构建及其免疫特性研究

Construction and Immunogenicity of a Recombinant Adenovirus Expressing the M Protein of Porcine Reproductive and Respiratory Syndrome Virus in Mice

本研究通过RT-PCR方法扩增猪繁殖与呼吸综合征病毒(PRRSV)S1株的M蛋白基因,将其克隆重组到人 血清5型腺病毒载体中,转染293细胞,制备重组腺病毒rAd-M。RT-PCR和IFA方法鉴定,结果表明rAd-M可表 达M基因的mRNA和M蛋白。纯化的rAd-M重组腺病毒经293细胞连续传25代,滴度稳定为107.8 TCID50／ mL。动物免疫试验结果表明,该重组腺病毒rAd-M能够刺激机体产生PRRSV的特异性抗体免疫和细胞免疫应 答反应,从而为PRRSV结构蛋白功能及其基因工程疫苗研究奠定了基础。

To screen out vaccine candidate against PRRSV, adenovirus vector was used to express the M protein of Porcine reproductive and respiratory syndrome virus （PRRSV）. A recombinant adenovirus was constructed and the expression of the M protein was identified by RT-PCR and indirect immunofluorescence assay （IFA）. The purified recombinant adenovirus M （rAd-M） was passaged 25 times in 293A cells. The titer of stocks of rAd-M was stably 10^7.8／CID50/mL. Furthermore, the recombinant M protein adenovirus could induce PRRSV specific humoral immunity and cell mediated immunity in mice. It indicated that the recombinant adenovirus expressing the main structural proteins of PRRSV is a potentially viable candidate vaccine against PRRSV.