摘要
为构建质粒稳定型山羊痘DNA疫苗,采用PCR与限制性酶切技术去除真核表达质粒pcDNA3.1(+)的氨苄抗性bla基因启动子序列,构建改良质粒pmcDNA3.1(+),然后插入山羊痘病毒P32基因,获得重组表达质粒pmcDNA3.1-P32,通过TSS法将其转化至减毒沙门氏菌中,构建成功携带山羊痘DNA疫苗的重组减毒沙门氏菌SL7207(pmcDNA3.1-P32);体内和体外试验结果表明,重组质粒pmcDNA3.1-P32在沙门氏菌中的稳定性显著高于pcDNA3.1-P32。这为下一步减毒沙门氏菌介导的山羊痘DNA免疫研究奠定了基础。
In order to construct the goatpox DNA vaccine of stable plasmid,the methods of PCR and restriction enzyme were applied to remove the promoter sequence of ampicillin resistance gene（bla gene） in plasmid pcDNA3.1（＋） to construct the pmcDNA3.1（＋）.The P32 gene of goatpox virus was subcloned into eukaryotic expression vector pmcDNA3.1（＋）.The recombinant plasmid was transformed into attenuated Salmonella typhimurium SL7207,and the recombinant was designated as SL7207（pmcDNA3.1-P32）.In vitro and in vivo experiments showed that the plasmid stability of pmcDNA3.1-P32 was apparently higher than that of pcDNA3.1-P32 in SL7207.It provided the foundation for the future research of mucosal immunity with attenuated Salmonella typhimurium.
出处
《生物技术通报》
CAS
CSCD
北大核心
2011年第3期105-110,共6页
Biotechnology Bulletin
基金
贵州省优秀科技教育人才省长专项基金项目(黔省专合[2006]12号)
国家自然科学基金主任基金项目(30940054)
关键词
减毒沙门氏菌
稳定
山羊痘
疫苗
Attenuated Salmonella typhimurium Stability Goatpox Vaccine
