摘要
用PCR扩增了MBP—ZOT质粒上的ZOT基因，该基因编码264～399位氨基酸残基C末端的15ku △G片段，并与经改造后的质粒P^BCX、连接。将重组后的质粒PMLAG在大肠杆菌BL21中表达，将表达的目的蛋白纯化，再将p^BCX-△G片段与传染性腔上囊炎病毒VP2包涵体蛋白通过滴鼻免疫途径对14日龄SPF鸡进行联合免疫试验，分别于免疫后第10、20d采集血清样品，进行ELISA试验，检测免疫后IBD抗体效价。动物免疫试验发现：融合蛋白P^BCX △G起到了明显免疫增强效果，与VP2重组蛋白联合免疫2次后，血清IBD的VP2 IgG抗体效价是单独用VP2免疫组的3倍，明显高于空白免疫组。
The carboxyl-terminal fragment of approximately 15 ku of zonula occludens toxin（ZOT） gene encoding the 264-399 amino acid residue was amplified by PCR and then cloned into the prokaryotic expression plasmids p^BCX. The active protein expressed in Escherichia coli was purified. The inclusion body from VP2 gene of infectious bursal disease vir＇us（IBDV） was used to intranasally vaccinate the 14-day-old SPF chickens, combined with purified protein P^BCX △G, with a booster 10 days later. The serum samples were collected every 10 days after the last immunization. ELISA were used to detect the IBD antibodies titer of anti-VP2. The results showed that the fusion protein P^BCX △G exhibited an adjuvant activity. The IBD antibody titer of VP2 in the chickens immunized with VP2 in the presence P^BCX △G was approxi- mately 3-fold higher than those of the control group after the second immunization.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第2期135-139,共5页
Chinese Veterinary Science
基金
广东省农业攻关项目(2002B21404)
关键词
霍乱弧菌
封闭带毒素
原核表达
传染性腔上囊炎
免疫
Vibrio cholerae
zonula occludens toxin
prokaryotic expression
infectious bursal disease
immunization
