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 Publication | VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
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ISSN | 0165-2427 |
Volume | 121Issue:1-2Pages:44-57 |
Abstract | 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.; 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. |
Subtype | Article |
Keyword | Traf1 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 Area | Immunology ; Veterinary Sciences |
DOI | 10.1016/j.vetimm.2007.08.001 |
WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
Indexed By | SCI |
Language | 英语 |
WOS Research Area | Immunology ; Veterinary Sciences |
WOS Subject | Immunology ; Veterinary Sciences |
WOS ID | WOS:000252817300005 |
WOS Keyword | NECROSIS-FACTOR RECEPTOR ; NF-KAPPA-B ; SIGNAL-TRANSDUCING PROTEINS ; VIRUS TRANSFORMING PROTEIN ; FACTOR FAMILY ; CYTOPLASMIC DOMAIN ; ADAPTER PROTEINS ; TERMINAL KINASE ; FACTOR (TRAF)-1 ; ACTIVATION |
Citation statistics | |
Document Type | 期刊论文 |
Identifier | http://ir.ihb.ac.cn/handle/152342/8282 |
Collection | 期刊论文 |
Corresponding Author | Nie, P, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China |
Affiliation | 1.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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