摘要
【目的】构建捻转血矛线虫DNA疫苗,并研究该疫苗对山羊的免疫保护效果。【方法】将捻转血矛线虫重要保护性抗原H11的编码基因分H11-1、H11-2和H11-3三部分,分别亚克隆到基因疫苗载体pcDNA4/HisMaxC中,用酶切反应、序列测定等方法鉴定重组疫苗。将纯化的DNA疫苗肌肉注射山羊后,用RT-PCR、Westernblotting和ELISA方法检测疫苗在山羊体内的转录、翻译和诱导IgG的产生。第2次免疫后2W用10000条捻转血矛线虫第3期幼虫攻击实验动物,检测山羊虫卵排出、虫卵孵化率、成虫数量等免疫保护性指标。【结果】获得了捻转血矛线虫H11DNA疫苗,疫苗第1次免疫7d后在山羊肌肉组织中进行了转录,第2次免疫7d后,DNA疫苗获得了翻译,并诱导机体产生了相应的抗体。与对照组比较,试验组山羊排出虫卵减少58.8%、虫卵孵化率减少75.3%、成虫减少68.2%。【结论】构建了捻转血矛线虫H11DNA疫苗,该DNA疫苗对山羊具有较好的免疫保护效果。
【Objective】The study was carried out to construct DNA vaccines for Haemonchus contortus, and test the protective effects in goats. 【Method】Three gene fragments encoding parts of H. contortus H11, H11-1, H11-2 and H11-3, were inserted into pcDNA4/His Max C. The recombinants were identified by double restriction enzyme digestion and sequence analysis. Lambs were immunized with DNA vaccines about 28 and 14 days before challenged by 10 000 H. contortus L3. The expression of vaccines in goats was analyzed by RT-PCR and Western blotting. While the IgG stimulated with the vaccines was checked by ELISA. After challenge with L3, EPG (eggs per gram feces), L3 hatched from eggs and worm burdens of lambs were counted. 【Result】 Transcriptions and translations of the vaccines in goat muscles were confirmed at seven days post primary immunization and one week after boosting. And the serum anti-H11 antibodies were inspected by ELISA about ten days after the first immunization. After challenged with L3, it was found that EPG, percentage of L3 hatched from eggs and worm burdens of lambs, immunized with H11 DNA vaccines reduced by 58.8%, 75.3% and 68.2%, respectively, compared with the goats administered with control pcDNA4/His Max C. 【Conclusion】 The results showed that the recombinant plasmids carrying H11 gene could protect lambs from the infection of H. contortus effectively.
出处
《中国农业科学》
CAS
CSCD
北大核心
2007年第12期2869-2875,共7页
Scientia Agricultura Sinica
基金
国家自然基金(30371078)
江苏省自然基金(BK2006145)
