摘要
为构建鸡柔嫩艾美耳球虫(E.tenella)重组真核表达质粒并检测其免疫保护性,本实验将E.tenella表面抗原基因3-1E(E)与鸡IFN-γ、IL-2基因分别串联在真核表达载体pcDNA3.1(+)中,构建真核重组表达质粒pcDNA-E-IFNγ、pcDNA-E-IL2,同时构建对照质粒pcDNA-E、pcDNA-IFNγ、pcDNA-IL2。经酶切鉴定得到预期大小目的片段,测序结果与预期序列一致。将构建的重组质粒免疫鸡只,经western blot检测表明该重组质粒在鸡体内可以正常表达;对感染鸡只的免疫保护效果显示,pcDNA-E-IFNγ、pcDNA-E-IL2组抗球虫指数(ACI)分别为181.04、187.30,具有高效的抗球虫效果,为将其进一步研制成为具有实用价值的抗球虫疫苗奠定了基础。
To evaluate the protection of recombinant plasmid co-expressing Eimeria tenella 3-1E protein and chicken IFN-γ or IL-2 against E.tenella,the 3-1E gene was fused with chicken IFN-γ and IL-2 genes to construct eukaryotic expression recombinant plasmid pcDNA-E-IFNγ and pcDNA-E-IL2,respectively.The chickens were inoculated with the recombinant plasmids at dosage of 100 μg per chicken via leg muscle injections twice in interval of 7 days and challenged with 5×104 E.tenella sporulated oocysts at 7 days post immunization.The results indicated that the recombinant plasmids expressed in chickens detected by western blot and the anticoccidial index（ACI） of pcDNA-E-IFNγ and pcDNA-E-IL2 was 181.04 and 187.30,respectively.The results showed that the plasmids had high anticoccidial effect and provided basis for further development of anticoccidial vaccine.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2011年第11期902-905,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家十一五科技支撑计划重大项目(2006BAD06A08)
哈尔滨市科技创新人才研究专项(2006RFQXN016)
