禽多杀性巴氏杆菌C_(48-1)成熟外膜蛋白H重组亚单位疫苗免疫效果的研究

Studies on immune efficacy of recombinant subunit vaccine with mature outer membrane protein H against avian Pasteurella multocida C_(48-1)

将融合表达禽多杀性巴氏杆菌成熟外膜蛋白H(OmpmH)的重组菌pGEX-ompmH／ BL21大量培养,在最佳诱导条件下诱导表达,表达产物经蛋白酶剪切及亲和层析纯化,得到Ompm H基因的原核表达产物,将其与弗氏完全佐剂混合制成油乳剂亚单位疫苗,用该疫苗肌肉注射接种5周龄鸡,首免后每周采血检测抗体,二免后第2周用10 LD50禽多杀性巴氏杆菌强毒菌株C48-1 进行攻击。结果显示,OmpmH具有良好的免疫原性,能诱导鸡体产生特异性抗体,可抵抗强毒菌株C48-1的致死性攻击,免疫效果优于禽多杀性巴氏杆菌弱毒疫苗。

In order to detect immune efficacy of a recombinant subunit vaccine with mature outer membrane protein H against avian Pasteurella multocida C48-1, recombinant plasmid pGEX-ompm H was cultured in E. coli BL21 and induced under the best condition. After analysis of solubility of the expressed product, the target protein Ompm H was purified with PreScissionTMPrOtease and Glutathion SepharoseTM 4B. Five-weeks-old chickens were inoculated intramuscularly with the purified protein. Each week sera were collected from chickens to examine antibodies after first immunization. The immunized chickens were challenged with 10LD50 avian P. multocida C48-1 two weeks-after second immunization. Results showed that the mature outer membrane protein H had better immunogenicity, and could induce the immune response efficiently. The immune effect of the recombinant subunit vaccine was better than that of the attenuated live vaccine, because some chickens vaccinated with the emulsified recombinant subunit vaccine were protected while the unvaccinated chickens in the control experienced 100% of mortality following challenge.