摘要
将堆型艾美耳球虫保定株裂殖子3-1E基因,插入到真核表达载体pcDNA3.1(+)中,构建重组表达质粒pcDNA-3-1E,用碱裂解法大量提取重组表达质粒,纯化后免疫接种1周龄雏鸡。分别设高(100μg)、中(50μg)、低(25μg)3个剂量组,同时设活卵囊接种组、非免疫感染组和健康对照组。在7日龄时首免,14日龄加强免疫,1周后用2.5×105个E.acervulina保定株球虫孢子化卵囊进行攻毒试验。结果显示50μg的剂量可使试验鸡产生较强的抗球虫效果,攻虫鸡的卵囊产量下降67.67%,肠道病变记分降低66.96%,并使鸡的相对增重率达88.36%,ACI值为176.06。
The DNA recombinant plasmid pcDNA3-3-1E was constructed by inserting ORF sequence of 3-1E gene of Baoding strain Eimeria acervulina merozoite into eukaryotic vector pcDNA3.l（＋）.The resulting recombinant plasmids were extracted by Plasmid Large-scale Extraction Kit（centrifugation columniation type,TIANGEN Beijing Biotechnology Co.,Ltd.） and purified.A-week-chickens were immunized with the purified recombinant expression plasmid pcDNA-3-3-1E with dose of 100,50,25 μg per bird,respectively,and strengthened immunity one week later.Simultaneously,chickens immunized with E.acervulina oocysts,nonimmunized infected chickens,healthy chicken were used as control.The chickens were inoculated with 2.5×105 sporulated E.acervulina oocysts（Baoding strain） at a week post immunization.The immuno protection of recombinant expression plasmid pcDNA3-3-1E of Eimeria acervulina for chicken was observed.The results showed that the test chickens with dose of 50μg plasmid pcDNA-3-3-1E could produce stronger anticoccodial effect.It＇s oocyst output reduced 67.67 %,intestinal lesion score fell 66.96%,and the relative weight of chicken rate was 88.36%,ACI value was 176.06.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2011年第8期1138-1141,共4页
Chinese Journal of Veterinary Science
基金
河北省自然科学基金资助项目(C2007000523)
国家自然科学基金资助项目(30571394)
