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学科主题: Fisheries; Immunology; Marine & Freshwater Biology; Veterinary Sciences
题名: Molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, Epinephelus coioides
作者: Wei, SN ; Huang, YH ; Cai, J ; Huang, XH ; Fu, J ; Qin, QW
通讯作者: qinqw@scsio.ac.cn
关键词: Epinephelus coioides ; c-type lysozyme ; Antimicrobial ; Antivirus
刊名: FISH & SHELLFISH IMMUNOLOGY
发表日期: 2012
卷: 33, 期:2, 页:186-196
部门归属: [Wei, Shina; Huang, Youhua; Cai, Jia; Huang, Xiaohong; Fu, Jing; Qin, Qiwei] Chinese Acad Sci, S China Sea Inst Oceanol, Key Lab Marine Bioresources Sustainable Utilizat, Guangzhou 510301, Guangdong, Peoples R China; [Wei, Shina; Cai, Jia; Fu, Jing] Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
项目归属: LMB
摘要: Lysozymes are key proteins of the host innate immune system against pathogen infection. In this study, a c-type lysozyme gene (Ec-lysC) was cloned and characterized from orange-spotted grouper, Epinephelus coioides. The full-length Ec-lysC cDNA is composed of 533 bp and encodes a polypeptide of 144-residue protein with 94% identity to lysC of Kelp grouper, Epinephelus bruneus. The genomic DNA of Ec-lysC consists of 4 exons and 3 introns, with a total length of 1897 bp. Amino acid sequence alignment showed that Ec-lysC possessed conserved catalytic residues (Glu50 and Asp67) and "GSTDYGIFQINS" motif. RT-PCR results showed that Ec-lysC transcript was most abundant in head kidney and less in muscle. The expression of Ec-lysC was differentially up-regulated in head kidney after stimulation with lipopolysaccharide (LPS), Vibrio alginolyticus and Singapore grouper iridovirus (SGIV). Subcellular localization analysis revealed that Ec-lysC was distributed predominantly in the cytoplasm. The recombinant Ec-lysC (rEc-lysC) had lytic activities against Gram-positive bacteria Micrococcus lysodeikticus, Staphylococcus aureus, Streptococcus iniae and Gram-negative bacteria V. alginolyticus. The lysozyme acted on M. lysodeikticus cell walls as shown by scanning electron microscopy (SEM). Furthermore, overexpression of Ec-lysC in grouper cells delayed the occurrence of CPE induced by SGIV and inhibited the viral gene transcription significantly. Taken together, Ec-lysC might play an important role in grouper innate immune responses to invasion of bacterial and viral pathogens. C-type lysozyme gene from E. coioides (Ec-lysC) was identified and characterized. (C) 2012 Elsevier Ltd. All rights reserved.
语种: 英语
WOS记录号: WOS:000306632800004
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.scsio.ac.cn/handle/344004/9852
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Wei, SN; Huang, YH; Cai, J; Huang, XH; Fu, J; Qin, QW.Molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, Epinephelus coioides,FISH & SHELLFISH IMMUNOLOGY,2012,33(2):186-196
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