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学科主题: Fisheries; Immunology; Marine & Freshwater Biology; Veterinary Sciences
题名: Structure, organization and expression of common carp (Cyprinus carpio L.) NKEF-B gene
作者: Huang, Rong1, 3; Gao, Long-Ying2; Wang, Ya-Ping1; Hu, Wei1; Guo, Qiong-Lin1
通讯作者: Guo, QL, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
关键词: NKEF-B ; NKEF-A ; NK cells ; Molecular cloning ; Common carp
刊名: FISH & SHELLFISH IMMUNOLOGY
发表日期: 2009-02-01
DOI: 10.1016/j.fsi.2008.10.013
卷: 26, 期:2, 页:220-229
收录类别: SCI
文章类型: Article
部门归属: [Huang, Rong; Wang, Ya-Ping; Hu, Wei; Guo, Qiong-Lin] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; [Gao, Long-Ying] Shenzhen Exit & Entry Inspect & Quarantine Bur, Key Lab Aquat Anim Dis, Shenzhen 518001, Peoples R China; [Huang, Rong] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
资助者: National Natural Science Foundation of China [30671599, 30871912]; National Basic Research Program of China [2009CB118704]; State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences [2006FB12]
类目[WOS]: Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
研究领域[WOS]: Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
摘要: Natural killer (NK) cell enhancing factor (NKEF) belongs to the newly defined peroxiredoxin (Prx) family. Its functions are to enhance NK cell cytotoxicity and to protect DNA and proteins from oxidative damage. In this study, a partial cDNA sequence of carp NKEF-B was isolated from thymus cDNA library. Subsequently, the full-length cDNA of carp NKEF-B was obtained by means of 3' and 5' RACE, respectively. The full-length cDNA of carp NKEF-B was 1022 bp, consisting of a 73 bp 5'-terminal untranslated region (UTR), a 355 bp T-terminal UTR, and a 594 bp open reading frame coding for a protein of 197 amino acids. Carp NKEF-B contained two consensus Val-Cys-Pro (VCP) motifs and three consensus cysteine (Cys-51, Cys-70 and Cys-172) residues. Sequence comparison showed that the deduced amino acid sequence of carp NKEF-B had an overall similarity of 74-96% to that of other species homologues. Phylogenetic analysis revealed that carp NKEF-B forms a cluster with other known teleost NKEF-Bs. Then, by PCR we obtained a 5.1 -k long genomic DNA of carp NKEF-B containing six exons and five introns. Realtime RT-PCR results showed that carp NKEF-B gene was predominantly detected in kidney and head kidney under un-infected conditions. Whereas under SVCV-infection condition, the expression of NKEF-B gene was significantly increased in blood cells, gill, intestine and spleen, but maintained in liver, and decreased significantly in kidney and head kidney. Finally, the rNKEF-B was constructed and expressed in Escherichia coli. By using an antibody against carp rNKEF-B, immunohistochemical study further indicated that NKEF-B positive cells are mainly some RBCs and a few epithelial cells in gill and intestine, and that under SVCV-infection condition, these positive cells or positive products in their cytoplasm were mainly increased in gill and spleen sections of carp. The results obtained in the present study will help to understand the function of NKEF-B in teleost innate immunity. (C) 2008 Elsevier Ltd. All rights reserved.
英文摘要: Natural killer (NK) cell enhancing factor (NKEF) belongs to the newly defined peroxiredoxin (Prx) family. Its functions are to enhance NK cell cytotoxicity and to protect DNA and proteins from oxidative damage. In this study, a partial cDNA sequence of carp NKEF-B was isolated from thymus cDNA library. Subsequently, the full-length cDNA of carp NKEF-B was obtained by means of 3' and 5' RACE, respectively. The full-length cDNA of carp NKEF-B was 1022 bp, consisting of a 73 bp 5'-terminal untranslated region (UTR), a 355 bp T-terminal UTR, and a 594 bp open reading frame coding for a protein of 197 amino acids. Carp NKEF-B contained two consensus Val-Cys-Pro (VCP) motifs and three consensus cysteine (Cys-51, Cys-70 and Cys-172) residues. Sequence comparison showed that the deduced amino acid sequence of carp NKEF-B had an overall similarity of 74-96% to that of other species homologues. Phylogenetic analysis revealed that carp NKEF-B forms a cluster with other known teleost NKEF-Bs. Then, by PCR we obtained a 5.1 -k long genomic DNA of carp NKEF-B containing six exons and five introns. Realtime RT-PCR results showed that carp NKEF-B gene was predominantly detected in kidney and head kidney under un-infected conditions. Whereas under SVCV-infection condition, the expression of NKEF-B gene was significantly increased in blood cells, gill, intestine and spleen, but maintained in liver, and decreased significantly in kidney and head kidney. Finally, the rNKEF-B was constructed and expressed in Escherichia coli. By using an antibody against carp rNKEF-B, immunohistochemical study further indicated that NKEF-B positive cells are mainly some RBCs and a few epithelial cells in gill and intestine, and that under SVCV-infection condition, these positive cells or positive products in their cytoplasm were mainly increased in gill and spleen sections of carp. The results obtained in the present study will help to understand the function of NKEF-B in teleost innate immunity. (C) 2008 Elsevier Ltd. All rights reserved.
关键词[WOS]: KILLER-ENHANCING FACTOR ; TROUT ONCORHYNCHUS-MYKISS ; TRANSCRIPTION FACTORS ; CHANNEL CATFISH ; HYDROGEN-PEROXIDE ; SPRING VIREMIA ; CELLS ; IDENTIFICATION ; CLONING ; PEROXIREDOXINS
语种: 英语
WOS记录号: WOS:000264329800003
ISSN号: 1050-4648
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/7816
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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作者单位: 1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2.Shenzhen Exit & Entry Inspect & Quarantine Bur, Key Lab Aquat Anim Dis, Shenzhen 518001, Peoples R China
3.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China

Recommended Citation:
Huang, Rong; Gao, Long-Ying; Wang, Ya-Ping; Hu, Wei; Guo, Qiong-Lin.Structure, organization and expression of common carp (Cyprinus carpio L.) NKEF-B gene,FISH & SHELLFISH IMMUNOLOGY,2009,26(2):220-229
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