Molecular cloning and characterization of a new G-type lysozyme gene (Ec-lysG) in orange-spotted grouper, Epinephelus coioides
Wei, SN; Huang, YH; Huang, XH; Cai, J; Wei, JG; Li, PF; Ouyang, ZL; Qin, QW; qinqw@scsio.ac.cn
2014
发表期刊DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
ISSN0145-305X
卷号46期号:2页码:401-412
摘要Lysozyme acts as an innate immunity molecule against pathogen infection. In this study, a new G-type lysozyme gene with a typical G-type lysozyme domain (designated as Ec-lysG) was cloned and characterized from the orange-spotted grouper, Epinephelus coioides. The full-length Ec-lysG cDNA contains 1419 bp and encodes a 256-residue protein containing a 25-residue signal peptide at the N-terminus. BLAST analysis reveals Ec-lysG shares 64% identity with Siniperca chuatsi, but 63% to another reported G-type lysozyme from orange-spotted grouper (OSG-lysG). The genomic DNA of Ec-lysG contains four exons and three introns, with a total length of 2062 bp. An amino acid sequence alignment showed that Ec-lysG shares the fundamental structural features of G-type lysozyme, including the catalytic residues, substrate binding sites, and soluble lytic transglycosylase domain. Quantitative PCR showed that Ec-lysG transcript is most abundant in the head kidney, and less abundant in the heart. The expression of Ec-lysG was differentially upregulated in the head kidney after stimulation with lipopolysaccharide, Vibrio alginolyticus, and Singapore grouper iridovirus (SGIV). A subcellular localization analysis showed that Ec-lysG is distributed predominantly in the cytoplasm. Recombinant Ec-lysG (rEc-lysG) has optimal activity at pH 7.5 and 35 degrees C. rEc-lysG showed lytic activities against Gram-positive bacterium Streptococcus iniae, Staphylococcus aureus, and Micrococcus lysodeikticus, and the Gram-negative bacterium V. alginolyticus. Scanning electron microscopy (SEM) showed that rEc-lysG acts on M. lysodeikticus cell walls. The overexpression of Ec-lysG in grouper cells did not significantly delay the occurrence of the cytopathic effect (CPE) induced by SGIV, and did not inhibit viral gene transcription. In conclusion, Ec-lysG might be a potent antibacterial protein, with a role in innate immunity. (C) 2014 Elsevier Ltd. All rights reserved.
部门归属[Wei, Shina ; Huang, Youhua ; Huang, Xiaohong ; Cai, Jia ; Wei, Jingguang ; Li, Pengfei ; Ouyang, Zhengliang ; Qin, Qiwei] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangzhou 510301, Guangdong, Peoples R China ; [Cai, Jia] Guangdong Ocean Univ, Coll Fishery, Zhanjiang 524088, Peoples R China ; [Li, Pengfei] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
关键词Epinephelus Coioides G-type Lysozyme Antimicrobial Activity Innate Immunity
学科领域Immunology ; Zoology
资助者National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203] ; National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203] ; National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203] ; National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203]
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收录类别sci
资助项目LMB
资助者National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203] ; National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203] ; National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203] ; National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203]
WOS记录号WOS:000340216600031
引用统计
被引频次:14[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/10384
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,Huang, XH,et al. Molecular cloning and characterization of a new G-type lysozyme gene (Ec-lysG) in orange-spotted grouper, Epinephelus coioides[J]. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,2014,46(2):401-412.
APA Wei, SN.,Huang, YH.,Huang, XH.,Cai, J.,Wei, JG.,...&qinqw@scsio.ac.cn.(2014).Molecular cloning and characterization of a new G-type lysozyme gene (Ec-lysG) in orange-spotted grouper, Epinephelus coioides.DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,46(2),401-412.
MLA Wei, SN,et al."Molecular cloning and characterization of a new G-type lysozyme gene (Ec-lysG) in orange-spotted grouper, Epinephelus coioides".DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 46.2(2014):401-412.
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