Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 13 2 2021 04 10 235 242 EN Ehsan Zamani Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran Jamil Zargan Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran Hossein Honari Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran Abbas Hajizade Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran Ashkan Haji Noor Mohammadi Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran Hani Keshavarz Alikhani Department of Biology, Razi University, Kermanshah, Iran Ahmad Heidari Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Zanjan, Zanjan, Iran Mohammad Hossein Pour Department of Biological Sciences, Faculty of Sciences, Imam Hossein University, Tehran, Iran 2020 05 16 2021 02 22 Background and Objectives: Aspergillus clavatus antimicrobial peptide (AcAMP) is a fungi-derived peptide with a broad spectrum of activity against pathogenic bacteria and fungi. Natural antimicrobial peptides, including AcAMP, have attracted many attentions in the development of new natural antibiotics against pathogenic bacteria, especially multidrug resistant ones. Materials and Methods: In the present study, acamp gene was codon-optimized and chemically synthesized in pUC57 cloning vector, subcloned into pET28a (+) expression vector and transferred into competent Escherichia coli BL21 (DE3) cells. The expression of AcAMP was induced by addition of Isopropyl β- d-1-thiogalactopyranoside (IPTG) and the expressed peptide was purified by Ni-NTA. BALB/c mice were immunized with the purified peptide and the ability of the immunized mice sera for the detection of the native AcAMP secreted by A. clavatus IRAN 142C was examined through ELISA and Western blotting techniques. Results: Both ELISA and Western blotting demonstrated the ability of the sera of the immunized mice to detect the native AcAMP. Conclusion: The results of the present work show that the raised antibody against recombinant AcAMP can be used to detect AcAMP peptide, an issue which paves the way to develop detection kits for the detection of AcAMP-producing organisms, purification of this valuable peptide for further investigations. https://ijm.tums.ac.ir/index.php/ijm/article/view/2597 https://ijm.tums.ac.ir/index.php/ijm/article/download/2597/1338