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en Heterologous Expression and Functional Characterization of CAP18 from Oryctolagus cuniculus زیست شناسی ملکولی Molecular Biology مقالات اصلی Original Article <div style="text-align: justify;">Background: Antimicrobial peptides belong to the innate defence system of creatures. These peptides attach to the bacterial membrane in order to die microorganisms by penetrating them. Hence, biotechnology researchers pay more attention to produce antimicrobial peptides for use in various fields. The studies showed that rabbit tissue with inflammation and skin ulcers would be producing CAP18 peptide, which belongs to the cathelicidin group. Methods: In this study, the optimized sequence of the cap18 gene was placed into the pPICZAα plasmid after the alpha-factor signal and transformed into Pichia pastoris (X-33 strain). Purification of the recombinant peptide was done based on its histidine tail at C-terminal, and western blotting method was used to demonstrate the purification of rCAP18. The antibacterial activity of the purified and desalted rCAP18 was investigated at different concentrations against pathogenic bacteria. Results: The maximum expression level of rCAP18 (17.5 kDa) was seen 90 h after induction of alcohol oxidase I (AOX1) promoter with methanol. The concentration of rCAP18 was 33 mg/L after purification with Ni-NTA Sepharose column. The function of rCAP18 (4.3, 5.7, 7 μg/ml) was investigated against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Results showed that %CFU/cm2 reached 28% after P. aeruginosa cells treatment with 7 μg/ml of rCAP18. Conclusions: This study presented the findings related to heterologous expression of cap18 gene, and evaluation of rCAP18 antibacterial effects. Our results showed that rCAP18 plays a significant role in inhibiting bacterial growth, especially Gram-negative bacteria.</div> Antibacterial activity, Cathelicidin, Recombinant CAP18, Heterologous expression. 622 632 http://rbmb.net/browse.php?a_code=A-10-733-1&slc_lang=en&sid=1 Mahla Nikpoor 100319475328460012456 100319475328460012456 No Department of Agricultural Biotechnology, Shahid Bahonar University of Kerman, Kerman, Iran. Azadeh Lohrasbi-Nejad a.lohrasbi@uk.ac.ir 100319475328460012457 100319475328460012457 Yes Department of Agricultural Biotechnology, Shahid Bahonar University of Kerman, Kerman, Iran. Jafar Zolala 100319475328460012458 100319475328460012458 No Department of Agricultural Biotechnology, Shahid Bahonar University of Kerman, Kerman, Iran.