Volume 10, Issue 4 (Vol.10 No.4 Jan 2022)                   rbmb.net 2022, 10(4): 622-632 | Back to browse issues page

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Nikpoor M, Lohrasbi-Nejad A, Zolala J. Heterologous Expression and Functional Characterization of CAP18 from Oryctolagus cuniculus. rbmb.net. 2022; 10 (4) :622-632
URL: http://rbmb.net/article-1-679-en.html
Department of Agricultural Biotechnology, Shahid Bahonar University of Kerman, Kerman, Iran.
Abstract:   (736 Views)
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.
Full-Text [PDF 364 kb]   (367 Downloads)    
Type of Article: Original Article | Subject: Molecular Biology
Received: 2021/03/30 | Accepted: 2021/04/6 | Published: 2022/02/7

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