为在大肠杆菌中高效表达出可溶性的传染性造血器官坏死病病毒（IHNV）基质蛋白M，并分析其免疫原性，构建了pMAL-c2x-m原核表达质粒，优化表达条件并纯化MBP-M融合蛋白；对蛋白进行Western blot和DDA鉴定；将鉴定后的蛋白免疫BALB/c小鼠，利用Western blot鉴定鼠抗MBP-M血清，通过间接ELISA检测该血清效价。结果显示：在大肠杆菌中获得了可溶性的MBP-M蛋白，其相对分子质量约为64 kDa，与预期结果一致；MBP-M蛋白在IPTG为0.5 mmol/L，25 ℃诱导12 h时上清表达量较高；经Western blot鉴定，MBP-M蛋白可与鼠抗血清发生特异性反应，而不与阴性对照血清发生反应；ELISA检测血清效价达1:25 600。结果表明，本研究获得的MBP-M蛋白具有较好的免疫原性，这为IHNV基因工程疫苗研发奠定了基础。
In order to express soluble matrix protein M of infectious hematopoietic necrosis virus （IHNV-M）from Escherichia coli（E. coli）and to analyze its immunogenicity，a prokaryotic expression plasmid was constructed and named as pMAL-c2x-m，the expression conditions was optimized and then MBP-M fusion protein was purified. The purified protein was identified by Western blot and DDA，and then was used to immunize BALB/c mice，and the mouse antiserum was identified by Western blot，whose titer was determined by indirect ELISA. The results showed that soluble MBP-M protein was obtained from E. coli，and its relative molecular mass was about 64 kDa as expected；the expression level of MBP-M protein in supernatant was higher when IPTG was 0.5 mmol/L and the bacteria solution was induced at 25 ℃ for 12 h；as identified by Western blot，MBP-M protein could specifically react with mouse antiserum，but failed with negative control serum；and the titer of serum antibodies reached 1:25 600 by indirect ELISA. In conclusion，the immunogenicity of the obtained MBP-M protein was good，which laid a foundation for future researches and development of genetic engineering vaccines of IHNV.