Society of African Journal Editors

African Journal of Microbiology Research

Cloning and expression of C-terminal of Clostridium perfringens type A enterotoxin and its biological activity

Authors:

Atieyeh Taherian Fard1, Fariha Hasan1, Mojgan Bandehpour2, Nariman Mosaffa2, Fatemeh Mashhadi Abbas3, Abdul Hameed1, Aamer Ali Shah1 and Bahram Kazemi2*

Journal: African Journal of Microbiology Research

Pathogenic clostridia produce exocellular toxins that resemble lipoteichoic acid and are described as super antigens. These toxins stimulate T-cell receptor-carrying lymphocytes in peripheral blood and have been used to study immunodeficiency diseases and cancers. The CPE C-terminal region from one of the local type A strain was cloned in the pET32a vector its expression induced with IPTG. The expressed protein was purified by Ni-NTA affinity chromatography and tested for biological activity with Vero cells assay. This region of Clostridium perfringens enterotoxin (CPE) has a predominant ligand-binding activity. In the present study, the biological activity of the C-terminal region of local purified CPE came under study with Vero cell assay, guinea pig skin test and mouse test to evaluate for future use as a therapeutic purpose. The result of this study showed that, the study’s local purified C-CPE had cytotoxic activity in Vero cells even at the minimum dilution of 0.625 ng after a 4-h incubation period. It caused transient increase in capillary permeability in guinea pigs. C-CPE did not have systemic effect on Balb/c mice. The use of the C-CPE peptide may provide a novel way to target drugs to Claudine-expressing cells.

 

Key words: Cloning, gene expression, Clostridium perfringens enterotoxin (CPE), vero cells, nigrosin, guinea pig skin test, mouse test, Claudine.