Molecular cloning and characterization of a new G-type lysozyme gene (Ec-lysG) in orange-spotted grouper, Epinephelus coioides

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	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
刊名	DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
	2014
卷号	46 期号:2 页码:401-412
关键词	Epinephelus coioides G-type lysozyme Antimicrobial activity Innate immunity
ISSN号	0145-305X
通讯作者	qinqw@scsio.ac.cn
中文摘要	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.
学科主题	Immunology ; Zoology
收录类别	SCI
资助信息	National Basic Research Program of China (973) [2012CB114402]; National High Technology Development Program of China (863) [2014AA093507, 2012AA092201]; Chinese Academy of Sciences [XDA11020203]
原文出处	ELSEVIER SCI LTD
WOS记录号	WOS:000340216600031
公开日期	2014-12-11
内容类型	期刊论文
源URL	[http://ir.scsio.ac.cn/handle/344004/10384]
专题	南海海洋研究所_中科院海洋生物资源可持续利用重点实验室
推荐引用方式 GB/T 7714	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.,...&Qin, QW.(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.