中国畜牧兽医2012，Vol.39Issue(3)：36-41,6.
D型产气荚膜梭菌ε毒素基因表达及其免疫保护作用的初步研究
Expression of Epsilon-toxin Gene of Clostridium perfringens Type D and its Primary Immunological Protective Function
摘要
Abstract
Epsilon-toxin (etoxin) gene was amplified from chromosomal DNA of Clostridium perfringens type D (C60-2) by polymerase chain reaction (PCR) .and a 906 bp epsilon toxin gene fragment was obtained. Sequence analysis indicated that the homology of the nucleotide sequence of the strain to those other reference strains was more than 99%. The expression plas-mid pET32a-ETX was constructed by inserting the epsilon toxin gene into the prokaryotic expression vector pET32a. The plas-mid pET32a-ETX was transformated into E. coli BL21 (DE3) plys and the recombinant strain BL21 (pET32a-ETX) was obtained. Then,the transformants were induced to express with IPTG. The specific 54 ku protein was detected by SDS-PAGE and the immunogenicity of the expressed epsilon toxin was confirmed by Western blotting and ELISA. The obtained recombinant protein was transformed into epsilon toxoid vaccine by adding 0. I% formaldehyde into epsilon toxin. The protective immune response was proved after the mice was immunized with epsilon toxoid vaccine. The results showed that the recombinan-ted strain BL21(pET32a-ETX) could be as a candidate of epsilon toxoid vaccine to provide protective immune response against C. Perfringens type D infection.关键词
D型产气荚膜梭菌/ε毒素/类毒素/免疫保护Key words
Clostridium perfringens type D/epsilon-toxin/ toxoid jimmunoprotection分类
农业科技引用本文复制引用
王光华,蔺国珍,郑福英,曹小安,宫晓炜,周继章,邱昌庆..D型产气荚膜梭菌ε毒素基因表达及其免疫保护作用的初步研究[J].中国畜牧兽医,2012,39(3):36-41,6.基金项目
国家科技基础性工作专项(2008FY210200). （2008FY210200）
