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Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter
Jiang, Jun; Zhang, Yi-Bing; Li, Shun; Yu, Fei-Fei; Sun, Fan; Gui, Jian-Fang; Zhang, YB, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China
2009-09-01
Source PublicationMOLECULAR IMMUNOLOGY
ISSN0161-5890
Volume46Issue:15Pages:3131-3140
AbstractGrass carp hemorrhagic virus (GCHV)-induced gene 2 (Gig2) is a novel gene previously identified from UV-inactivated GCHV-treated Carassius auratus blastulae embryonic (CAB) cells, suggesting that it should play a pivotal role in the interferon (IFN) antiviral response. In this study, a polyclonal anti-Gig2 antiserum was generated and used to study the inductive expression pattern by Western blot analysis, showing no basal expression in normal CAB cells but a significant up-regulation upon UV-inactivated GCHV, polyinosinic:polycytidylic acid (Poly I:Q and recombinant IFN (rIFN). However, constitutive expression of Gig2 is observed in all tested tissues from grass carp (Ctenopharyngodon idellus), and Poly I:C injection increases the relative amount of Gig2 protein in skin, spleen, trunk kidney, gill, hindgut and thymus. Moreover, the genomic sequence covering the whole Gig2 ORF and the upstream promoter region were amplified by genomic walking. Significantly, the Gig2 promoter contains three IFN-stimulated response elements (ISREs), nine GAAA/TfTC motifs and five gamma-IFN activating sites (GAS), which are the characteristics of genes responsive to both type I IFN and type 11 IFN. Subsequently, the complete Gig2 promoter sequence was cloned into pGL3-Basic vector, and its activity was measured by luciferase assays in the transfected CAB cells. The Gig2 promoter-driven construct is highly induced in CAB cells after treatment with Poly I:C or rIFN, and the functional capability is dependent on IFN regulatory factor 7 (IRF7), because its activity can be stimulated by IRF7. Collectively, the data provide strong evidence that Gig2 is indeed a novel IFN inducible gene and its expression is likely dependent on IRF7 upon Poly I:C or IFN. (C) 2009 Elsevier Ltd. All rights reserved.; Grass carp hemorrhagic virus (GCHV)-induced gene 2 (Gig2) is a novel gene previously identified from UV-inactivated GCHV-treated Carassius auratus blastulae embryonic (CAB) cells, suggesting that it should play a pivotal role in the interferon (IFN) antiviral response. In this study, a polyclonal anti-Gig2 antiserum was generated and used to study the inductive expression pattern by Western blot analysis, showing no basal expression in normal CAB cells but a significant up-regulation upon UV-inactivated GCHV, polyinosinic:polycytidylic acid (Poly I:Q and recombinant IFN (rIFN). However, constitutive expression of Gig2 is observed in all tested tissues from grass carp (Ctenopharyngodon idellus), and Poly I:C injection increases the relative amount of Gig2 protein in skin, spleen, trunk kidney, gill, hindgut and thymus. Moreover, the genomic sequence covering the whole Gig2 ORF and the upstream promoter region were amplified by genomic walking. Significantly, the Gig2 promoter contains three IFN-stimulated response elements (ISREs), nine GAAA/TfTC motifs and five gamma-IFN activating sites (GAS), which are the characteristics of genes responsive to both type I IFN and type 11 IFN. Subsequently, the complete Gig2 promoter sequence was cloned into pGL3-Basic vector, and its activity was measured by luciferase assays in the transfected CAB cells. The Gig2 promoter-driven construct is highly induced in CAB cells after treatment with Poly I:C or rIFN, and the functional capability is dependent on IFN regulatory factor 7 (IRF7), because its activity can be stimulated by IRF7. Collectively, the data provide strong evidence that Gig2 is indeed a novel IFN inducible gene and its expression is likely dependent on IRF7 upon Poly I:C or IFN. (C) 2009 Elsevier Ltd. All rights reserved.
SubtypeArticle
KeywordGig2 Interferon Interferon-stimulated Gene (Isg) Irf7 Poly i:c Promoter Interferon-stimulated Response Elements Expression Regulation
Department[Jiang, Jun; Zhang, Yi-Bing; Li, Shun; Yu, Fei-Fei; Sun, Fan; Gui, Jian-Fang] Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Subject AreaBiochemistry & Molecular Biology ; Immunology
DOI10.1016/j.molimm.2009.05.183
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding Organization973 National Basic Research Program of China [2004CB117403]; 863 High Technology Research Program of China [2007AA09Z423]; National Natural Science Foundation of China [30671617, 30871922]; Innovation Project of Institute of Hydrobiology, Chinese Academy of Sciences [085A01-1-301] ; 973 National Basic Research Program of China [2004CB117403]; 863 High Technology Research Program of China [2007AA09Z423]; National Natural Science Foundation of China [30671617, 30871922]; Innovation Project of Institute of Hydrobiology, Chinese Academy of Sciences [085A01-1-301] ; 973 National Basic Research Program of China [2004CB117403]; 863 High Technology Research Program of China [2007AA09Z423]; National Natural Science Foundation of China [30671617, 30871922]; Innovation Project of Institute of Hydrobiology, Chinese Academy of Sciences [085A01-1-301] ; 973 National Basic Research Program of China [2004CB117403]; 863 High Technology Research Program of China [2007AA09Z423]; National Natural Science Foundation of China [30671617, 30871922]; Innovation Project of Institute of Hydrobiology, Chinese Academy of Sciences [085A01-1-301]
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Immunology
WOS SubjectBiochemistry & Molecular Biology ; Immunology
WOS IDWOS:000270489800029
WOS KeywordTROUT ONCORHYNCHUS-MYKISS ; NF-KAPPA-B ; CARP-HEMORRHAGE-VIRUS ; RAINBOW-TROUT ; ATLANTIC SALMON ; PARALICHTHYS-OLIVACEUS ; CAB CELLS ; MX GENE ; MOLECULAR CHARACTERIZATION ; JAPANESE FLOUNDER
Funding Organization973 National Basic Research Program of China [2004CB117403]; 863 High Technology Research Program of China [2007AA09Z423]; National Natural Science Foundation of China [30671617, 30871922]; Innovation Project of Institute of Hydrobiology, Chinese Academy of Sciences [085A01-1-301] ; 973 National Basic Research Program of China [2004CB117403]; 863 High Technology Research Program of China [2007AA09Z423]; National Natural Science Foundation of China [30671617, 30871922]; Innovation Project of Institute of Hydrobiology, Chinese Academy of Sciences [085A01-1-301] ; 973 National Basic Research Program of China [2004CB117403]; 863 High Technology Research Program of China [2007AA09Z423]; National Natural Science Foundation of China [30671617, 30871922]; Innovation Project of Institute of Hydrobiology, Chinese Academy of Sciences [085A01-1-301] ; 973 National Basic Research Program of China [2004CB117403]; 863 High Technology Research Program of China [2007AA09Z423]; National Natural Science Foundation of China [30671617, 30871922]; Innovation Project of Institute of Hydrobiology, Chinese Academy of Sciences [085A01-1-301]
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Cited Times:27[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/7564
Collection期刊论文
Corresponding AuthorZhang, YB, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China
AffiliationChinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Jiang, Jun,Zhang, Yi-Bing,Li, Shun,et al. Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter[J]. MOLECULAR IMMUNOLOGY,2009,46(15):3131-3140.
APA Jiang, Jun.,Zhang, Yi-Bing.,Li, Shun.,Yu, Fei-Fei.,Sun, Fan.,...&Zhang, YB, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China.(2009).Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter.MOLECULAR IMMUNOLOGY,46(15),3131-3140.
MLA Jiang, Jun,et al."Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter".MOLECULAR IMMUNOLOGY 46.15(2009):3131-3140.
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