Molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, Epinephelus coioides
Wei, SN; Huang, YH; Cai, J; Huang, XH; Fu, J; Qin, QW; qinqw@scsio.ac.cn
2012
发表期刊FISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
卷号33期号:2页码:186-196
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.
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
关键词Epinephelus Coioides C-type Lysozyme Antimicrobial Antivirus
学科领域Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
语种英语
资助项目LMB
WOS记录号WOS:000306632800004
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文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/9852
专题中科院海洋生物资源可持续利用重点实验室
通讯作者qinqw@scsio.ac.cn
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Wei, SN,Huang, YH,Cai, J,et al. Molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, Epinephelus coioides[J]. FISH & SHELLFISH IMMUNOLOGY,2012,33(2):186-196.
APA Wei, SN.,Huang, YH.,Cai, J.,Huang, XH.,Fu, J.,...&qinqw@scsio.ac.cn.(2012).Molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, Epinephelus coioides.FISH & SHELLFISH IMMUNOLOGY,33(2),186-196.
MLA Wei, SN,et al."Molecular cloning and characterization of c-type lysozyme gene in orange-spotted grouper, Epinephelus coioides".FISH & SHELLFISH IMMUNOLOGY 33.2(2012):186-196.
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