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Characterization and expression analysis of TNFR-associated factor 1 (TRAF1) in grass carp Ctenopharyngodon idella
Xu, Z. Y.1,2,3; Sun, B. J.1,2; Chang, M. X.1,2; Nie, P.1,2; Nie, P, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2008-01-15
Source PublicationVETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
ISSN0165-2427
Volume121Issue:1-2Pages:44-57
AbstractTNF receptor associated factor 1 (TRAF1) plays an important role in regulating the TNF signaling and protecting cells from apoptosis. In the present study, a TRAF1 gene has been cloned from grass carp (Ctenopharyngodon idella) by reverse transcription (RT)-PCR and rapid amplification of cDNA ends (RACE). The full-length cDNA is 2235 bp, including a 250 bp 5' UTR (untranslated region), a 1659 bp open reading frame, and a 326 bp 3'UTR. The polyadenylation signal (AATAAA, AATAA) and one mRNA instability motif (AUUUA) were found followed by a poly (A) tail in the 3'UTR. No signal peptide or transmembrane region has been found in the putative amino acids of grass carp TRAF1 (gcTRAF1). The putative amino acids of gcTRAF1 share 72% identity with the homologue in zebrafish. It is characterized by a zinc finger at the N-terminus and a TRAF domain (contains one TRAF-C and one TRAF-N) at the C-terminus. The identity of the TRAF domain among all the TRAF1 homologues in vertebrates varies from 52% to 58%, while the identities of TRAF-C were almost the same as 70%. The recombinant gcTRAF1 has been constructed successfully and expressed in Escherichia coli by using pET-32a expression vector. The polyclonal antibody for rabbit has been successfully obtained. The expression of gcTRAF1 in different organs was examined by real-time quantitative PCR and Western blotting, respectively. It was widely distributed in heart, head kidney, thymus, brain, gill, liver, spleen, and trunk kidney. This is the first report of TRAF1 homologue molecule found in fish. (c) 2007 Elsevier B.V. All rights reserved.; TNF receptor associated factor 1 (TRAF1) plays an important role in regulating the TNF signaling and protecting cells from apoptosis. In the present study, a TRAF1 gene has been cloned from grass carp (Ctenopharyngodon idella) by reverse transcription (RT)-PCR and rapid amplification of cDNA ends (RACE). The full-length cDNA is 2235 bp, including a 250 bp 5' UTR (untranslated region), a 1659 bp open reading frame, and a 326 bp 3'UTR. The polyadenylation signal (AATAAA, AATAA) and one mRNA instability motif (AUUUA) were found followed by a poly (A) tail in the 3'UTR. No signal peptide or transmembrane region has been found in the putative amino acids of grass carp TRAF1 (gcTRAF1). The putative amino acids of gcTRAF1 share 72% identity with the homologue in zebrafish. It is characterized by a zinc finger at the N-terminus and a TRAF domain (contains one TRAF-C and one TRAF-N) at the C-terminus. The identity of the TRAF domain among all the TRAF1 homologues in vertebrates varies from 52% to 58%, while the identities of TRAF-C were almost the same as 70%. The recombinant gcTRAF1 has been constructed successfully and expressed in Escherichia coli by using pET-32a expression vector. The polyclonal antibody for rabbit has been successfully obtained. The expression of gcTRAF1 in different organs was examined by real-time quantitative PCR and Western blotting, respectively. It was widely distributed in heart, head kidney, thymus, brain, gill, liver, spleen, and trunk kidney. This is the first report of TRAF1 homologue molecule found in fish. (c) 2007 Elsevier B.V. All rights reserved.
SubtypeArticle
KeywordTraf1 Grass Carp Cdna Expression Ctenopharyngodon Idella
Department[Xu, Z. Y.; Sun, B. J.; Chang, M. X.; Nie, P.] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; [Xu, Z. Y.; Sun, B. J.; Chang, M. X.; Nie, P.] Chinese Acad Sci, Inst Hydrobiol, Lab Fish Dis, Wuhan 430072, Peoples R China; [Xu, Z. Y.] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Subject AreaImmunology ; Veterinary Sciences
DOI10.1016/j.vetimm.2007.08.001
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaImmunology ; Veterinary Sciences
WOS SubjectImmunology ; Veterinary Sciences
WOS IDWOS:000252817300005
WOS KeywordNECROSIS-FACTOR RECEPTOR ; NF-KAPPA-B ; SIGNAL-TRANSDUCING PROTEINS ; VIRUS TRANSFORMING PROTEIN ; FACTOR FAMILY ; CYTOPLASMIC DOMAIN ; ADAPTER PROTEINS ; TERMINAL KINASE ; FACTOR (TRAF)-1 ; ACTIVATION
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Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8282
Collection期刊论文
Corresponding AuthorNie, P, 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, Inst Hydrobiol, Lab Fish Dis, Wuhan 430072, Peoples R China
3.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Recommended Citation
GB/T 7714
Xu, Z. Y.,Sun, B. J.,Chang, M. X.,et al. Characterization and expression analysis of TNFR-associated factor 1 (TRAF1) in grass carp Ctenopharyngodon idella[J]. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY,2008,121(1-2):44-57.
APA Xu, Z. Y.,Sun, B. J.,Chang, M. X.,Nie, P.,&Nie, P, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(2008).Characterization and expression analysis of TNFR-associated factor 1 (TRAF1) in grass carp Ctenopharyngodon idella.VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY,121(1-2),44-57.
MLA Xu, Z. Y.,et al."Characterization and expression analysis of TNFR-associated factor 1 (TRAF1) in grass carp Ctenopharyngodon idella".VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY 121.1-2(2008):44-57.
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