Please use this identifier to cite or link to this item: http://archive.cmb.ac.lk:8080/xmlui/handle/70130/5334
Title: Genomic characterization and expression profiles upon bacterial infection of a novel cystatin B homologue from disk abalone ( Haliotis discus discus)
Authors: Premachandra, H. K. A.
Wan, Q.
Elvitigala, D. A. S
De Zoysa, M.
Choi, C. Y.
Whang, I.
Lee, J.
Keywords: Cysteine protease inhibitors
Cystatin B
Disk abalone
Bacterial challenge
Papain inhibition Transcriptional analysis
Issue Date: 2012
Publisher: Elsevier
Citation: Premachandra, H. K. A., Wan, Q., Elvitigala, D. A. S., De Zoysa, M., Choi, C. Y., Whang, I., & Lee, J. (2012). Genomic characterization and expression profiles upon bacterial infection of a novel cystatin B homologue from disk abalone (Haliotis discus discus). Developmental & Comparative Immunology, 38(4), 495-504.
Abstract: Cystatins are a large family of cysteine proteinase inhibitors which are involved in diverse biological and pathological processes. In the present study, we identified a gene related to cystatin superfamily, AbCyt B, from disk abalone Haliotis discus discus by expressed sequence tag (EST) analysis and BAC library screening. The complete cDNA sequence of AbCyt B is comprised of 1967 nucleotides with a 306 bp open reading frame (ORF) encoding for 101 amino acids. The amino acid sequence consists of a single cystatinlike domain, which has a cysteine proteinase inhibitor signature, a conserved Gly in N-terminal region, QVVAG motif and a variant of PW motif. No signal peptide, disulfide bonds or carbohydrate side chains were identified. Analysis of deduced amino acid sequence revealed that AbCyt B shares up to 44.7% identity and 65.7% similarity with the cystatin B genes from other organisms. The genomic sequence of AbCyt B is approximately 8.4 Kb, consisting of three exons and two introns. Phylogenetic tree analysis showed that AbCyt B was closely related to the cystatin B from pacific oyster (Crassostrea gigas) under the family 1.Functional analysis of recombinant AbCyt B protein exhibited inhibitory activity against the papain, with almost 84% inhibition at a concentration of 3.5 lmol/L. In tissue expression analysis, AbCyt B transcripts were expressed abundantly in the hemocyte, gill, mantle, and digestive tract, while weakly in muscle, testis, and hepatopancreas. After the immune challenge with Vibrio parahemolyticus, the AbCyt B showed significant (P < 0.05) up-regulation of relative mRNA expression in gill and hemocytes at 24 and 6 h of post infection, respectively. These results collectively suggest that AbCyst B is a potent inhibitor of cysteine proteinases and is also potentially involved in immune responses against invading bacterial pathogens in abalone.
URI: http://archive.cmb.ac.lk:8080/xmlui/handle/70130/5334
Appears in Collections:Department of Basic Sciences & Social Sciences

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