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Molecular cloning, genomic structure and antimicrobial activity of PtALF7, a unique isoform of anti-lipopolysaccharide factor from the swimming crab Portunus trituberculatus
Liu, Yuan1; Cui, Zhaoxia1; Li, Xihong1,2; Song, Chengwen1,2; Shi, Guohui1,2; Wang, Chunlin3; Cui, ZX
2013-02-01
发表期刊FISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
卷号34期号:2页码:652-659
文章类型Article
摘要Anti-lipopolysaccharide factors (ALFs), as the potent antimicrobial peptides, are becoming predominant candidates for potential therapeutic agents of bacterial and viral diseases. In this study, a unique isoform of ALF (PtALF7) was identified from hemocytes cDNA library of the swimming crab Portunus trituberculatus. The PtALF7 cDNA contained an open reading frame (ORF) of 372 bp encoding 123 amino acids. The deduced peptide of PtALF7 shared high similarity with our previously reported PtALF1-3 but low with PtALF4-6. The PtALF7 gene consisted of three exons interrupted by two introns, and was clearly transcribed from different genomic loci compared with other PtALF isoforms. Totally 128 SNPs including 12 in coding region and 116 in noncoding region were detected in PtALF7 gene by direct sequencing of 20 samples. The mRNA expression of PtALF7 transcript was primarily observed in hemocytes followed by gill and eyestalk, but barely detectable in hepatopancreas. After challenge with Vibrio alginolyticus, a main pathogen causing high mortality in P. trituberculatus, the PtALF7 transcript in hemocytes showed a clear time-dependent response expression pattern with obvious decrease at 6 h and significant increase at 24 h. The recombinant PtALF7 protein exhibited antimicrobial activity against the test Gram-negative and Gram-positive bacteria, but did not inhibit the growth of fungus Pichia pastoris. These results together indicate a potential involvement for PtALF7 in the innate immune response of P. trituberculatus. (C) 2012 Elsevier Ltd. All rights reserved.; Anti-lipopolysaccharide factors (ALFs), as the potent antimicrobial peptides, are becoming predominant candidates for potential therapeutic agents of bacterial and viral diseases. In this study, a unique isoform of ALF (PtALF7) was identified from hemocytes cDNA library of the swimming crab Portunus trituberculatus. The PtALF7 cDNA contained an open reading frame (ORF) of 372 bp encoding 123 amino acids. The deduced peptide of PtALF7 shared high similarity with our previously reported PtALF1-3 but low with PtALF4-6. The PtALF7 gene consisted of three exons interrupted by two introns, and was clearly transcribed from different genomic loci compared with other PtALF isoforms. Totally 128 SNPs including 12 in coding region and 116 in noncoding region were detected in PtALF7 gene by direct sequencing of 20 samples. The mRNA expression of PtALF7 transcript was primarily observed in hemocytes followed by gill and eyestalk, but barely detectable in hepatopancreas. After challenge with Vibrio alginolyticus, a main pathogen causing high mortality in P. trituberculatus, the PtALF7 transcript in hemocytes showed a clear time-dependent response expression pattern with obvious decrease at 6 h and significant increase at 24 h. The recombinant PtALF7 protein exhibited antimicrobial activity against the test Gram-negative and Gram-positive bacteria, but did not inhibit the growth of fungus Pichia pastoris. These results together indicate a potential involvement for PtALF7 in the innate immune response of P. trituberculatus. (C) 2012 Elsevier Ltd. All rights reserved.
关键词Portunus Trituberculatus Anti-lipopolysaccharide Factor Snp Antimicrobial Activity
学科领域Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
DOI10.1016/j.fsi.2012.12.002
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收录类别SCI
语种英语
WOS研究方向Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS类目Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS记录号WOS:000315076100028
WOS关键词ANTI-LPS FACTOR ; SHRIMP PENAEUS-MONODON ; BLACK TIGER SHRIMP ; ERIOCHEIR-SINENSIS ; MARSUPENAEUS-JAPONICUS ; SCYLLA-PARAMAMOSAIN ; CRYSTAL-STRUCTURE ; BINDING DOMAIN ; FACTOR ALF ; MUD CRAB
WOS标题词Science & Technology ; Life Sciences & Biomedicine
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被引频次:21[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/16606
专题海洋生态与环境科学重点实验室
实验海洋生物学重点实验室
通讯作者Cui, ZX
作者单位1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100039, Peoples R China
3.Ningbo Univ, Sch Marine Sci, Ningbo 315211, Zhejiang, Peoples R China
第一作者单位实验海洋生物学重点实验室
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Liu, Yuan,Cui, Zhaoxia,Li, Xihong,et al. Molecular cloning, genomic structure and antimicrobial activity of PtALF7, a unique isoform of anti-lipopolysaccharide factor from the swimming crab Portunus trituberculatus[J]. FISH & SHELLFISH IMMUNOLOGY,2013,34(2):652-659.
APA Liu, Yuan.,Cui, Zhaoxia.,Li, Xihong.,Song, Chengwen.,Shi, Guohui.,...&Cui, ZX.(2013).Molecular cloning, genomic structure and antimicrobial activity of PtALF7, a unique isoform of anti-lipopolysaccharide factor from the swimming crab Portunus trituberculatus.FISH & SHELLFISH IMMUNOLOGY,34(2),652-659.
MLA Liu, Yuan,et al."Molecular cloning, genomic structure and antimicrobial activity of PtALF7, a unique isoform of anti-lipopolysaccharide factor from the swimming crab Portunus trituberculatus".FISH & SHELLFISH IMMUNOLOGY 34.2(2013):652-659.
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