摘要
为研究副鸡禽杆菌外膜蛋白GcbG的免疫原性,克隆了副鸡禽杆菌荚膜蛋白合成相关的编码基因gcbG,成功构建了原核表达质粒pGEX-5X-gcbG,并将其转化到大肠杆菌BL21(DE3)菌株中进行诱导表达。结果表明,重组菌在28℃培养条件下经1mmol/L的IPTG诱导6h可分泌表达GcbG蛋白,且其表达量最高。Western-blot分析表明,所表达的蛋白能与副鸡禽杆菌阳性血清发生特异性免疫印迹反应,说明所表达的蛋白具有较好的反应原性。动物试验表明,所表达的蛋白能为鸡提供较好的保护力。上述试验结果为进一步研究荚膜蛋白的生物学特性及亚单位疫苗的开发奠定了基础。
To investigate the immunogenicity of Avibacterium paragallinarum outer membrane protein GcbG,the gcbGgene was cloned and transformend into Escherichia coli BL21（DE3）.The recombinant pGEX-5X-gcbG vector in E.coli BL21（DE3）was induced with 1mmol/L IPTG,and induced for 6hat28℃.Western-blot results showed that the protein has a specific reaction against A.paragallinarumpositive serum,which means that the protein had a good reactionogenicity with the positive serum.Animal test results indicated that the recombinant protein can protect SPF chicken from challenge.It appeared a foundation for further studies on biological characteristics of capsule protein and development of subunit vaccine.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2016年第1期58-63,共6页
Chinese Veterinary Science
基金
国家自然科学基金项目(31272558)
国家"十二五"高技术研究发展计划(863)项目(2011AA10A210)
北京市自然科学基金项目(6102009)
关键词
副鸡禽杆菌
gcbG基因
原核表达
动物试验
Avibacterium paragallinarum
gcbGgene
prokaryotic expression
animal experiment
