IHB OpenIR  > 鱼类生物学及渔业生物技术研究中心  > 期刊论文
Expression pattern, promoter activity and bactericidal property of beta-defensin from the mandarin fish Siniperca chuatsi
Wang, Gailing1,2,3; Li, Junhua1,2; Zou, Pengfei1,2; Xie, Haixia1; Huang, Bei1,2; Nie, Pin1; Chang, Mingxian1; Chang, MX (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.
2012-09-01
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume33Issue:3Pages:522-531
Abstractbeta-Defensin (BD) are cysteine-rich, cationic antimicrobial peptides which play an important role in innate immune system against invading microbes. In the present study, the cDNA cloning, expression analysis, transcriptional regulation and antimicrobial activity of beta-defensin (ScBD) from mandarin fish (Siniperca chuatsi) were characterized. The cDNA sequence of ScBD is 596 bp which encodes a protein of 63 amino acids (aa). The ScBD gene comprises three exons and two introns. The signal peptide is located in the first exon. ScBD contains 6 cysteines, and belongs to fish defensin 2 group based on phylogenetic analysis. Real-time quantitative PCR results showed that the mRNA transcripts of ScBD were distributed mainly in mucosal and lymphoid organs/tissues including intestine, gill, head kidney, kidney and spleen, with the highest level observed in spleen. Western blotting analysis revealed that the ScBD protein was abundant in head kidney, gill and spleen. A total of 3268 bp 5' flanking region of the ScBD gene promoter was sequenced, which contained a number of putative transcriptional binding sites for transcription factors. These transcription factors were analyzed using in vitro luciferase assay. The DNA region from position of -705 to -498 bp contains positive regulatory elements and that of -227 to +54 bp harbors the TATA which is essential for initiating gene expression. In addition, the ScBD peptide showed antibacterial activity against Escherichia call M15, Staphylococcus aureus and Aeromonas hydrophila, whilst no effect on Edwardsiella tarda. These data suggest that the ScBD is importantly involved in host immune responses to invasion of bacterial pathogens. (C) 2012 Elsevier Ltd. All rights reserved.; beta-Defensin (BD) are cysteine-rich, cationic antimicrobial peptides which play an important role in innate immune system against invading microbes. In the present study, the cDNA cloning, expression analysis, transcriptional regulation and antimicrobial activity of beta-defensin (ScBD) from mandarin fish (Siniperca chuatsi) were characterized. The cDNA sequence of ScBD is 596 bp which encodes a protein of 63 amino acids (aa). The ScBD gene comprises three exons and two introns. The signal peptide is located in the first exon. ScBD contains 6 cysteines, and belongs to fish defensin 2 group based on phylogenetic analysis. Real-time quantitative PCR results showed that the mRNA transcripts of ScBD were distributed mainly in mucosal and lymphoid organs/tissues including intestine, gill, head kidney, kidney and spleen, with the highest level observed in spleen. Western blotting analysis revealed that the ScBD protein was abundant in head kidney, gill and spleen. A total of 3268 bp 5' flanking region of the ScBD gene promoter was sequenced, which contained a number of putative transcriptional binding sites for transcription factors. These transcription factors were analyzed using in vitro luciferase assay. The DNA region from position of -705 to -498 bp contains positive regulatory elements and that of -227 to +54 bp harbors the TATA which is essential for initiating gene expression. In addition, the ScBD peptide showed antibacterial activity against Escherichia call M15, Staphylococcus aureus and Aeromonas hydrophila, whilst no effect on Edwardsiella tarda. These data suggest that the ScBD is importantly involved in host immune responses to invasion of bacterial pathogens. (C) 2012 Elsevier Ltd. All rights reserved.
SubtypeArticle
KeywordMandarin Fish Beta-defensin Gene Expression Promoter Activity Bactericidal Activity
Department[Wang, Gailing; Li, Junhua; Zou, Pengfei; Xie, Haixia; Huang, Bei; Nie, Pin; Chang, Mingxian] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; [Wang, Gailing; Li, Junhua; Zou, Pengfei; Huang, Bei] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China; [Wang, Gailing] Huanghuai Univ, Dept Bioengn, Zhumadian 463000, Henan, Peoples R China
DOI10.1016/j.fsi.2012.06.003
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationNational Natural Science Foundation of China [U0631010]; Government of Guangdong Province; National Basic Research Program of China (973 Program) [2009CB118703] ; National Natural Science Foundation of China [U0631010]; Government of Guangdong Province; National Basic Research Program of China (973 Program) [2009CB118703] ; National Natural Science Foundation of China [U0631010]; Government of Guangdong Province; National Basic Research Program of China (973 Program) [2009CB118703] ; National Natural Science Foundation of China [U0631010]; Government of Guangdong Province; National Basic Research Program of China (973 Program) [2009CB118703]
Indexed BySCI
Language英语
WOS Research AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS SubjectFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS IDWOS:000308057700007
WOS KeywordTROUT ONCORHYNCHUS-MYKISS ; NF-KAPPA-B ; ANTIMICROBIAL PEPTIDES ; FUNCTIONAL-CHARACTERIZATION ; ADAPTIVE IMMUNITY ; DENDRITIC CELLS ; HELA-CELLS ; INNATE ; GENE ; BETA-DEFENSIN-2
Funding OrganizationNational Natural Science Foundation of China [U0631010]; Government of Guangdong Province; National Basic Research Program of China (973 Program) [2009CB118703] ; National Natural Science Foundation of China [U0631010]; Government of Guangdong Province; National Basic Research Program of China (973 Program) [2009CB118703] ; National Natural Science Foundation of China [U0631010]; Government of Guangdong Province; National Basic Research Program of China (973 Program) [2009CB118703] ; National Natural Science Foundation of China [U0631010]; Government of Guangdong Province; National Basic Research Program of China (973 Program) [2009CB118703]
Citation statistics
Cited Times:22[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/17230
Collection鱼类生物学及渔业生物技术研究中心_期刊论文
Corresponding AuthorChang, MX (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.
Affiliation1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
3.Huanghuai Univ, Dept Bioengn, Zhumadian 463000, Henan, Peoples R China
Recommended Citation
GB/T 7714
Wang, Gailing,Li, Junhua,Zou, Pengfei,et al. Expression pattern, promoter activity and bactericidal property of beta-defensin from the mandarin fish Siniperca chuatsi[J]. FISH & SHELLFISH IMMUNOLOGY,2012,33(3):522-531.
APA Wang, Gailing.,Li, Junhua.,Zou, Pengfei.,Xie, Haixia.,Huang, Bei.,...&Chang, MX .(2012).Expression pattern, promoter activity and bactericidal property of beta-defensin from the mandarin fish Siniperca chuatsi.FISH & SHELLFISH IMMUNOLOGY,33(3),522-531.
MLA Wang, Gailing,et al."Expression pattern, promoter activity and bactericidal property of beta-defensin from the mandarin fish Siniperca chuatsi".FISH & SHELLFISH IMMUNOLOGY 33.3(2012):522-531.
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