Expression and identification of recombinant Clostridium difficile toxin B using Bacillus megaterium system / 中华传染病杂志
Chinese Journal of Infectious Diseases
; (12): 1-5, 2011.
Article
Dans Zh
| WPRIM
| ID: wpr-414212
Responsable en Bibliothèque :
WPRO
ABSTRACT
Objective To express and purify recombinant and biologically active Clostridium difficile toxin B (rTcdB). Methods The genes of TcdB were amplified by polymerase chain reaction (PCR) using chromosomal DNA from a toxigenic strain, and cloned into a shuttle vector pHis1522.The sequences of TcdB genes in the vector were verified by DNA sequencing. The construction was transformed into Bacillus megaterium protoplasts and the protein expression was driven by a xylose promoter. The purified protein was tested for biological activity. Results rTcdB was successfully purified from bacterial crude extracts. Approximately 5-10 mg of highly purified recombinant toxin was obtained from one liter of bacterial culture. The expressed rTcdB had molecular mass similar to the native toxin, and its biological activity was proved to be similar to its native counterpart after an extensive examination. Conclusion rTcdB with biological activities is successfully expressed in Bacillus megaterium.
Texte intégral:
1
Indice:
WPRIM
Type d'étude:
Diagnostic_studies
langue:
Zh
Texte intégral:
Chinese Journal of Infectious Diseases
Année:
2011
Type:
Article