猪繁殖与呼吸综合征病毒M蛋白基因疫苗的构建及在小鼠的免疫试验

Construction of Porcine Reproductive and Respiratory Syndrome Virus M Protein Gene Vaccine and Immunological Experiment on Mice

从PRRSV野毒株细胞培养上清提取RNA,用RT-PCR的方法扩增出M蛋白基因片段(525 bp),并将该基因克隆到pMD18-T载体,测序结果与PRRSV BJ-4等株同源性最接近.以pIRES-neo为载体,构建表达PRRSV M蛋白的基因疫苗,按100μg/只的DNA量免疫小鼠,二免后二周抗体水平显著高于同期免疫空载体pIRES-neo组,于免疫后14、28、42天后均能检测到较高的细胞免疫水平.这说明表达M蛋白的基因疫苗能够刺激机体产生免疫应答.

The membrane protein（M） gene of PRRSV was amplified by RT - PCR from the Marc - 145 cells infected with PRRSV street strain and inserted into the pMD18 - T vector, the homology of the sequence is the closest with the PRRSV BJ - 4 strain and other strains. The gene vaccine of expressing M protein from the PRRSV by pIRSEneo vector was Constructed and immunized the mice with a dosage of 100μg per mouse with the plasmids, the antibody level went up remarkably two weeks after the second injection of DNA vaccine and was higher than the empty vector group that immunized with pIRSEneo at the same time, and the cell - mediated immunity was detected at 14, 28 and 42 days from the mice immunized, it shows that the gene vaccine of expressing M protein to stimulate the mice in immune response.