Characterization of cathelicidin gene from buffalo (Bubalus bubalis)

Abstract

Wide spread and indiscriminate use of antibiotics is accompanied by the emergence of microorganism that are resistant to these agents. Therefore, new approaches are required to address the problem of antimicrobial resistance. Epithelial linings of living organisms are the source of various antimicrobial peptides. Keeping this in view, the present experiment was designed to characterize one of the important natural antimicrobial peptide gene, that is, cathelicidin gene from the buffalo (Bubalus bubalis) uterine epithelium and explored its potency for use as template for synthesis of novel antimicrobial agents. Total RNA was isolated from epithelial layer of buffalo uterus and reverse transcribed using designed primers. The amplified PCR product was purified and cloned. Positive clone was sequenced and result was analysed using laser gene software (DNA Star, USA). The cDNA of uterus cathelicidin had 516 bases with complete ORF from 6-452 bp. The predicted pre-propeptide comprised of 148 amino acids. Mature active peptide had 18 amino acids and had five each of arginine and tryptophan and four proline residues. From this study, it can be concluded that the buffalo (Bubalus bubalis) uterus expressed a potent antimicrobial peptide and amino acid sequence of mature peptide can be used as template for synthesis of novel antimicrobial agents.
 
Key words: Antimicrobial peptide, buffalo, cationic peptide and cathelicidin.