摘要
将携带鸡柔嫩艾美耳球虫5401基因的真核表达质粒pcDNA 3-5401的减毒沙门氏菌Z J111菌株(Z J111/pcD-NA 3-5401)口服接种于3日龄非免疫鸡,2周后进行第2次接种。结果表明,利用该减毒沙门氏菌作为载体具有相对安全性,用限制性酶切分析和PCR鉴定证实,体内试验和体外培养的重组质粒在受体菌Z J111菌株内比较稳定。用Z J111/pcDNA 3-5401(108cfu)口服接种非免疫鸡,2周后用相同剂量加强免疫1次,二免后2周用柔嫩艾美耳球虫孢子化卵囊攻击,观察其免疫效果。结果表明,重组Z J111/pcDNA 3-5401菌既能诱导产生抗鸡柔嫩艾美耳球虫抗体,也能显著增强淋巴细胞增殖水平(P<0.05);其抗球虫指数为164.98。试验结果显示,利用减毒沙门氏菌为载体传递DNA疫苗具有良好的安全性、稳定性和免疫原性。
The 3-day-old non-immune chicks were orally vaccinated with the recombinant attenuated Salmonella typhimuri- um strain ZJ111 carried pcDNA3-5401 expression plasmid encoding the fusion protein of Eimeria tenella and a booster was performed after 2 weeks. The recombinant plasmid pcDNA3-5401 was stable within the host strain in vitro and in vivo as shown by restriction enzymes analysis and PCR identification of the 5401 gene. In a vaccination test,3-day-old non-immune chicks were orally inoculated with ZJ111/pcDNA3-5401 with a dose of 108 cfu per chicks and boosted two weeks later. 10 chicks were challenged with a dose of 6×10^4 Eimeria tenella sporulated oocysts per bird at week 2 post boosting. The results showed that oral vaccination with ZJ111/pcDNA3-5401 induced strong humoral and celluar immune response and its anticoccidial index （ACI） was 164.98. The results indicate that the attenuated S. typhimurium as an oral delivery vector for DNA vaccine is of safety and stability and immunogenicity.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2006年第2期147-150,共4页
Chinese Journal of Veterinary Science
基金
杭州市科技局重点科研项目(2001112B14)
