Molecular cloning and functional characterizational of hepcidinfrom convict cichlid (Amatitlania nigrofasciata)

• Main Authors: Long-Si Liao, 廖龍喜
• Other Authors: Shao-Yang Hu
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/28218535039560312990

碩士 === 國立屏東科技大學 === 生物科技系所 === 101 === Hepcidin is an innate antimicrobial peptide, which existed widely in diverse animals and abound expressed in liver tissue to against foreign pathogens. The structure of hepcidin protein comprises multiple disulfide bonds and β-sheet motif. The bactericidal mechanism of hepcidin is depended on the osmotic stress due to the pore formation in cell membrane, which resulted from the interaction of positive-charged hepcidin and negative-charged cell membrane of pathogen. Ornamental fish is one of aquaculture industry and also face disease problems. In this study, hepcidin will be cloned from amatitlania nigrofasciata liver tissue and bactericidal activity will be also characterized. Full length of hepcidin gene had been cloned by degenerated primers, and complete hepcidin cDNA sequence of amatitlania nigrofasciata was composed 261 bases, and contained a translated region of 86 amino acids, which contained signal peptide, propeptide, and mature peptide. According to phylogenetic analysis, amatitlania nigrofasciata hepcidin is more similar to Oreochromis aureus hepcidin (80% identical). The result showed liver expreaaion of hepcidin in adult amatitlania nigrofasciata, and the expression of test was higher than other tissues. Minimal inhibitory concentration (MIC) values of synthetic peptide against the E.coli DH5α、Aermonas hydrophila、 Vibrio vulnificus and Streptococcus agalactine bacteria.