题目: Molecular cloning and characterization of two novel hepcidins from orange-spotted grouper, Epinephelus coioides 

作者: Zhou JG, Wei JG, Xu D, Cui HC, Yan Y, Ouyang ZL, Huang XH, Huang YH, Qin QW* 

刊物: Fish Shellfish Immunol. ，（IF=3.044，I区）

刊号: 2011, 30:559-568 

摘要: Orange-spotted grouper, Epinephelus coioides is one of the most important economic species of marine-cultured fish in China and Southeast Asia countries. However, very little information of the innate immune mechanisms against microbial pathogens is available in grouper, Epinephelus sp. Hepcidin, as an antimicrobial peptide (AMP), is a very important component in the innate immune system and widespread in fish. In this study, two novel types of hepcidin gene (designated EC-hepcidin1 and EC-hepcidin2) were cloned from E. coioides. They consist of open reading frames (ORFs) of 267 bp and 263 bp encoding the putative peptides of 88 and 87 amino acids, respectively. The homologous identity of deduced amino acid sequences between EC-hepcidin1 and EC-hepcidin2 is up to 79%, and predicted mature regions of both them have four cysteines residues. Genomic DNAs of both EC-hepcidin1 and EC-hepcidin2 consist of three exons and two introns. RT-PCR results showed that EC-hepcidin1 transcript was most abundant in liver and less in stomach. However, the transcript of EC-hepcidin2 was only detected in liver. The expressions of both EC-hepcidins were up-regulated by microbial and viral challenges, and iron overload, respectively, and EC-hepcidin1 was more responsive. The growth of Gram-negative bacterium of Vibrio vulnificus and Gram-positive bacterium of Staphylococcus aureus was inhibited by synthetic EC-hepcidins, and EC-hepcidin1 displayed stronger antimicrobial activity. The replication of Singapore grouper iridovirus (SGIV) was inhibited in the EC-hepcidin1 and EC-hepcidin2 over-expressed stable transfected fish cell lines (GS/pcDNA-Hep1, GS/pcDNA-Hep2) indicative of the antiviral activity of EC-hepcidins. These data should offer important information on the antimicrobial and antiviral roles of EC-hepcidins, and will be help to the better understanding of molecular mechanisms of grouper innate immunity. 

题目: A new leukocyte cell-derived chemotaxin-2 from marine fish grouper,Epinephelus coioides: Molecular cloning and expression analysis 

作者: Wei JG, Guo ML, Cui CH, Yan Y, Ouyang ZL, Qin QW* 

刊物: Fish Shellfish Immunol. ，（IF=3.044，I区）

刊号: 2011, 31:600-605 

摘要: Leukocyte cell-derived chemotaxin-2 (LECT2) is a multifunctional protein involved in cell growth, differentiation and autoimmunity. In this study, a new leukocyte cell-derived chemotaxin-2 (EcLECT2) gene was cloned from grouper, Epinephelus coioides, by rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA sequence of EcLECT2 was 595 bp in size, containing a 5′-untranslated region (UTR) of 44 bp and a 3′-UTR of 83 bp. The deduced protein sequence of the open reading frame (465 bp) showed highest similarity (81%) to the LECT2 of the fresh-water fish Larimichthys crocea. An abundant transcription of the determined EcLECT2 mRNA has been detected in liver and skin of grouper, E. coioides. Furthermore, the expression of EcLECT2 was differentially up-regulated in liver after infection with Staphyloccocus aureus,Vibrio vulnificus, Vibrio parahaemolyticus, Saccharomyces cerevisiae and Singapore grouper iridovirus (SGIV), while the expression was down-regulated after stimulation with Concanavalin A (Con A). Recombinant mature EcLECT2 (rEcLECT2) was successfully expressed in Escherichia coli BL21 (DE3), and the antiserum against EcLECT2 was obtained for further investigations. EcLECT2 may be an important molecule in the innate immunity of grouper. 

题目: Characterization of p38 MAPKs from orange-spotted grouper, Epinephelus coioides involved in SGIV infection

作者: Cai J, Huang YH, Wei SN, Huang XH, Ye FZ, Fu J, Qin QW*. 

刊物: Fish Shellfish Immunol. ，（IF=3.044，I区）

刊号: 2011, 31:1129-1136 

摘要: p38 mitogen-activated protein kinases (MAPKs) are broadly expressed signaling molecules that involves in the regulation of cellular responsible for various extracellular stimuli. In this study, three p38 MAPK genes (Ec-p38a, p38b and p38β) were cloned from grouper, Epinephelus coioides and their characteristics were investigated in vitro. Although Ec-p38a, p38b and p38β showed high homologies to other fish p38a MPAK, p38b MAPK and p38β MAPK, respectively, they all contained the conserved structures of Thr-Gly-Tyr (TGY) motif and substrate binding site Ala-Thr-Arg-Trp (ATRW). Phylogenetic analysis indicated that Ec-p38a, p38b and p38β are more closely related to those from fish than mammals. The tissue distribution patterns of Ec-p38a, p38b and p38β were different, and Ec-p38β was up-regulated most obviously in head kidney after Singapore grouper iridovirus (SGIV) infection. Overexpression of Ec-p38β in FHM cells delayed the occurrence of CPE induced by SGIV infection. Further analysis indicated that overexpression of Ec-p38β inhibited viral gene transcription and protein synthesis, as well as SGIV induced typical apoptosis in fish cells. Taken together, our data indicated that Ec-p38β played a crucial role in regulating apoptosis and virus replication during iridovirus infection.