A Streamlined Method to Obtain Biologically Active TcdA and TcdB Toxins from Clostridioides difficile.
Toxins (Basel)
; 16(1)2024 01 11.
Article
em En
| MEDLINE
| ID: mdl-38251254
ABSTRACT
The major virulence factors of Clostridioides difficile (C. difficile) are enterotoxins A (TcdA) and B (TcdB). The study of toxins is a crucial step in exploring the virulence of this pathogen. Currently, the toxin purification process is either laborious and time-consuming in C. difficile or performed in heterologous hosts. Therefore, we propose a streamlined method to obtain functional toxins in C. difficile. Two C. difficile strains were generated, each harboring a sequence encoding a His-tag at the 3' end of C. difficile 630∆erm tcdA or tcdB genes. Each toxin gene is expressed using the Ptet promoter, which is inducible by anhydro-tetracycline. The obtained purification yields were 0.28 mg and 0.1 mg per liter for rTcdA and rTcdB, respectively. In this study, we successfully developed a simple routine method that allows the production and purification of biologically active rTcdA and rTcdB toxins with similar activities compared to native toxins.
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Base de dados:
MEDLINE
Assunto principal:
Toxinas Bacterianas
/
Clostridioides difficile
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article