ISSN 2736-1756
Advanced Journal of Microbiology Research Vol. 2017
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2017 International Scholars Journals
Full Length Research Paper
Cloning and expression of C-terminal of Clostridium perfringens type A enterotoxin and its biological activity
Atieyeh Taherian Fard1, Fariha Hasan1, Mojgan Bandehpour 2, Nariman Mosaffa2, Fatemeh Mashhadi Abbas3, Abdul Hameed1, Aamer Ali Shah1 and Bahram Kazemi2*
1Department of Microbiology, Quaid - i -Azam University, Islamabad, Pakistan.
2Cellular and Molecular Biology Research Center, Shahid Beheshti University, M. C., Tehran, Iran.
3Department of Dentistry, Shahid Beheshti University, M. C, Tehran, Iran.
Accepted 5 April, 2017
Abstract
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.