摘要
构建表达细粒棘球蚴Eg95-Eg.ferritin融合蛋白的重组口服减毒鼠伤寒沙门菌活载体疫苗株并评价其生物学特性。将细粒棘球蚴Eg95-Eg.ferritin融合基因插入到表达载体pYA3341中,构建重组质粒pYA3341-Eg95-Eg.ferritin,将重组质粒分别电转入减毒鼠伤寒沙门菌X3770和X4550,获得重组疫苗菌株St-Eg95-Eg.ferritin,对重组菌的稳定性、生长状态及安全性进行分析。结果表明,经PCR和酶切鉴定成功构建重组质粒pYA3341-Eg95-Eg.ferritin,Western blot检测Eg95-Eg.ferritin蛋白在重组菌中获得表达;生物学特性研究结果表明,重组质粒可稳定存在,且不影响重组菌的生长状态,小鼠口服试验证实,重组菌安全无毒性。成功构建能稳定表达细粒棘球蚴Eg95-Eg.ferritin融合蛋白的口服减毒鼠伤寒沙门菌活载体疫苗株,将为研究包虫病口服基因工程疫苗奠定基础。
A recombinant attenuated Salmonella typhimuriuin oral live vector strain expressing the Eg 95-Eg. ferritin fusion antigen of Echinococcus granulosus was constructed and the biological characteristics of the recombinant vaccine strain were evaluated. The fusion gene Eg 95-Eg. ferritin was cloned and inserted into expression vector pYA3341. The recombinant plasmid pYA3341-Eg 95-Eg. ferritin was respectively electro-transformed into attenuated S. typhimurium vaccine strains X3730 and X4550 to construct the live vaccine strain St-Eg 95-Eg. ferritin. The vitro stability, growth characteristic and orally safety of recombinant vaccine St-Eg 95-Eg. ferritin were analyzed. The results showed that the recombinant plasmid pYA3341-Eg 95 Eg. ferritin was constructed successfully by enzyme digestion and PCR identification. The Eg 95 Eg. ferritin protein in the St-Eg 95-Eg. ferritin had a high level of expression, and the recombinant strain had a good genetic stability in vitro, growth states, safety, and non-toxic. Above studies suggested that the recombinant oral attenuated S. typhimurium St-Eg 95-Eg. ferritin was constructed successfully and could express stably Eg 95 Eg. ferritin protein. The results will provide a foundation for developing anti-Echinococcus granulosus oral genetic engineering vaccines.
出处
《西北农业学报》
CAS
CSCD
北大核心
2014年第10期44-48,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
宁夏自然科学基金(NZ12199)
宁夏卫生厅项目(2013125)
宁夏医科大学校级面上项目(XM2012032)
作者简介
第一作者：王志舁，男，博士，讲师，从事寄生虫分子免疫与疫苗研究。E-mail：wangzhisheng8239@163．com
通信作者：杨文，女，教授，主要从事临床血液学检验研究。E-mail：yangwen6211@163．com
