Design, construction, cloning and expression of beta Defensin and its effect on wound healing

authors:

avatar Nafiseh Karimi , avatar Behnaz Saffar , * , avatar Mohsen Mobini Dehkordi , avatar Kamran Ghaedi

Koomesh: Vol.17, issue 2; 358-365
published online: March 28, 2016
article type: Research Article
received: April 01, 2015
accepted: July 02, 2015

how to cite: Karimi N, Saffar B, Mobini Dehkordi M, Ghaedi K. Design, construction, cloning and expression of beta Defensin and its effect on wound healing. koomesh. 2016;17(2):e151353. 

Abstract

Introduction: Defensins are members of the largest family of antimicrobial peptides and due to their activity against bacteria, fungi, and viruses are very profitable as new antibiotics. The aim of this study was to design, synthesis, clone and expression of BNBD2 (Bovine Neutrophil Beta-Defensin2) gene to investigate the wound healing process. Materials and Methods: In this study, according to the preferred codon in E. coli, the BNBD2 gene was optimized and synthesized. The BNBD2 gene was sub cloned in vector pET-32a (+). The BNBD2 protein expression was assessed using Isopropyl-ß-D-thio-galactoside (IPTG) as an inducer and evaluating by SDS-PAGE. The potency of BNBD2 protein for healing was studied by creating a wound on a group of mice and treating them with BNBD2 protein in comparison with the control group. Results: The results showed that BNBD2 gene was successfully cloned into pET32a (+) vector. The expression of protein was induced by IPTG. There was a significant reduction in wound area in the treatment group in compare to the control group. Conclusiosn: Recombinant protein (BNBD2) was expressed successfully in prokaryotic system. This protein might be potentially beneficial for wound healing procedures in the future