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Development of porcine respiratory and reproductive syndrome virus replicon vector for foot-and-mouth disease vaccine

机译:猪口蹄疫疫苗的呼吸与生殖综合征病毒复制子的研制

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Purpose Foot-and-mouth disease (FMD) is an economically important global animal disease. To control FMD virus (FMDV) outbreaks, a lot of different novel approaches have been attempted. In this study, we proposed a novel porcine reproductive and respiratory syndrome virus (PRRSV) as a replicon vector to express FMDV structural protein. Materials and Methods PRRSV infectious clone (PRRSVK418DM) was used to develop an expression vector through the reverse genetic manipulation of PRRSV; FMDVP12A3C gene of serotype O was synthesized and used for an antigen. MARC-145 cells (African green monkey kidney epithelial cell line) were used for electroporation mediated transfection. The transfection or the expression of P12A3C and N protein of PRRSV was analyzed by either replicon containing PRRSV alone or by co-infection of helper PRRSV. Results We constructed PRRSVK418DM replicon vector containing FMDVP12A3C, and genome sequences were confirmed by subsequent sequence analysis. In vitro expression of P12A3C and PRRSV N protein was confirmed by immunofluorescence antibody assay using antibodies specific for PRRSV N protein (anti-PRRSV N MAb), FMDV-VP1 (anti-VP1 MAb). Conclusion The results indicate that PRRSV replicon vector can be a promising novel vector system to control FMDV and useful for vaccine development in the future.
机译:目的口蹄疫(FMD)是一种经济上重要的全球性动物疾病。为了控制口蹄疫病毒(FMDV)的爆发,尝试了许多不同的新颖方法。在这项研究中,我们提出了一种新型的猪繁殖与呼吸综合症病毒(PRRSV)作为表达FMDV结构蛋白的复制子载体。材料与方法使用PRRSV感染性克隆(PRRSVK418DM)通过PRRSV的反向遗传操作开发表达载体。合成了血清型O的FMDVP12A3C基因,并用于抗原。 MARC-145细胞(非洲绿猴肾上皮细胞系)用于电穿孔介导的转染。通过仅含有PRRSV的复制子或通过辅助PRRSV的共感染来分析PRRSV的P12A3C和N蛋白的转染或表达。结果我们构建了含有FMDVP12A3C的PRRSVK418DM复制子载体,并通过随后的序列分析证实了基因组序列。通过对PRRSV N蛋白(抗-PRRSV N MAb),FMDV-VP1(抗-VP1 MAb)具有特异性的抗体,通过免疫荧光抗体测定法证实了P12A3C和PRRSV N蛋白的体外表达。结论结果表明,PRRSV复制子载体可能是控制FMDV的有前途的新型载体系统,对将来的疫苗开发很有用。

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