基于SemlikiForest病毒RNA复制子的猪瘟RNA疫苗的初步研究

A Semliki Forest Virus RNA Replicon-based DNA Plasmid Encoding Classical Swine Fever Virus E2 Gene Induces Immune Response in Mice

将猪瘟病毒E2基因克隆于我们此前构建的衍生于Semliki Forest病毒(semliki forest virus,SFV)RNA复制子的新型真核表达载体pSFV1CS中,获得重组质粒pSFV1CS-E2。用纯化的pSFV1CS-E2分别转染BHK-21细胞和293T细胞,经间接免疫荧光试验检测显示,CSFV E2基因在转染细胞中得到表达。小鼠接种试验结果表明,10μg或100μg pSFV1CS-E2可诱导小鼠产生猪瘟特异性抗体。

The E2 gene of classical swine fever virus (CSFV) was cloned into the Bam HI/Sma I site of pSFV1CS, a eukaryotic expression vector derived from RNA replicon of Semliki Forest virus, creating a recombinant plasmid designated as pSFV1CS-E2. pSFV1CS-E2 was transfected into BHK-21 and 293T cells. The results showed that CSFV E2 was expressed efficiently in pSFV1CS-E2 transfected cells as demonstrated by indirect immunofluorescence assay (IFA) . Three of four BALB/c mice inoculated with 10 or 100 /μg of pSFV1CS-E2 developed E2-specific antibodies detected by an ELISA based on E. coli-expressed mutated E2 of CSFV. The recombinant plasmid expressing CSFV E2 gene can be used as a potential RNA vaccine to prevent CSFV infections.