摘要
为构建表达破伤风毒素C片段基因的减毒鼠伤寒沙门菌,将编码破伤风毒素C片段的基因插入到asd+的组成型表达载体pYA3333,转化asd基因突变的E.coliX6212,获得重组质粒pYA3333-TetC,再将重组质粒通过电穿孔法转入宿主菌,构建成表达破伤风毒素C片段基因的平衡致死的鼠伤寒沙门菌X4550(pYA3333-TetC)。免疫印迹试验证实,该重组菌表达的蛋白与预期的目的蛋白一致。重组菌X4550(pYA3333-TetC)的稳定性试验表明,在体外培养100代后,随机挑选的重组菌落全部都能生长,重组菌X4550(pYA3333-TetC)的生长曲线测定表明,X4550(pYA3333-TetC)和X4550(pYA3333)的生长状态基本一致。动物试验证明,该重组菌是安全的,破伤风毒素攻击后能提供良好的保护作用。
To construct a recombinant attenuated Salmonella typhimurium strain which can express tetanus toxin fragment C, the gene fragment encoding tetanus toxin fragment C was cloned into the expression vector pYA3333 containing asd gene, and a new recombinant plasmid pYA3333 TetC was generated. The recombinant plasmid pYA3333-TetC was transformed into an attenuated Salmonella typhimurium strain X4550 by electroporation, yielding the live vaccine strain X4550（pYA3333-TetC）, which was a balanced lethal recombinant. Stability and growth curve of the recombinant strain was determined. Results showed that the recombinant X4550（pYA3333-TetC） maintained good stability after 100 passages in vitro and the growth status of X4550 （pYA3333-TetC） was the same as that of X4550 （pYA3333）. The recombinant strain X4550（pYA3333-TetC） was safe and could provide significant protection against a lethal challenge with tetanus toxin in immunized mice.
出处
《中国兽医科技》
CSCD
北大核心
2005年第11期883-887,共5页
Chinese Journal of Veterinary Science and Technology
基金
国家自然科学基金项目(30425031)
全国优秀博士论文作者奖励计划项目(FANEDD200358)
关键词
破伤风毒素
鼠伤寒沙门菌
重组
tetanus toxin
Salmonella typhimurium
recombinant
