Cloning, characterization and expression analysis of SIMP (source of immunodominant MHC-associated peptides) in grass carp Ctenopharyngodon idella

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	Cloning, characterization and expression analysis of SIMP (source of immunodominant MHC-associated peptides) in grass carp Ctenopharyngodon idella
	Xu, Z. Y.1,2,3; Nie, P.1,2; Chang, M. X.1,2; Sun, B. J.1,2
刊名	FISH & SHELLFISH IMMUNOLOGY
	2008-06-01
卷号	24 期号:6 页码:701-714
关键词	SIMP grass carp cDNA gene organization promoter organ
ISSN号	1050-4648
通讯作者	Nie, P, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China
中文摘要	SIMP (source of immunodominant MHC-associated peptides) plays a key rote in N-linked glycosylation with the active site of oligosaccharyltransferase, being the source of MHC-peptides in the MHC I presentation pathway. In the present study, the SIMP gene has been cloned from grass carp Ctenopharyngodon idella by rapid amplification of cDNA ends (RACE). The full length of the cDNA sequence is 4384 bp, including a 1117 bp 5' UTR (untranslated region), a 2418 bp open reading frame, and a 849 bp 3' UTR. The deduced amino acids of the grass carp SIMP (gcSIMP) are a highly conserved protein with a STT3 domain and 11 transmembrane regions. The gcSIMP spans over more than 24,212 bp in length, containing 16 exons and 15 introns. Most encoding exons, except the first and the 15th, have the same length as those in human and mouse. The gcSIMP promoter contains many putative transcription factor binding sites, such as Oct-1, GCN4, YY1, Sp1, Palpha, TBP, GATA-1, C/EBP beta, and five C/EBP alpha binding sites. The mRNA expression of gcSIMP in different organs was examined by real-time PCR. The gcSIMP was distributed in all the organs examined, with the highest level in brain, followed by the level in the heart, liver, gill, trunk kidney, muscle, head kidney, thymus, and the lowest level in spleen. Furthermore, the recombinant gcSIMP has been constructed successfully and expressed in Escherichia coli by using pQE-40 vector, and the polyclonal antibody for rabbit has been successfully obtained, which was verified to be specific. Identification of gcSIMP will help to explore the function in fish innate immunity. (c) 2007 Elsevier Ltd. All rights reserved.
英文摘要	SIMP (source of immunodominant MHC-associated peptides) plays a key rote in N-linked glycosylation with the active site of oligosaccharyltransferase, being the source of MHC-peptides in the MHC I presentation pathway. In the present study, the SIMP gene has been cloned from grass carp Ctenopharyngodon idella by rapid amplification of cDNA ends (RACE). The full length of the cDNA sequence is 4384 bp, including a 1117 bp 5' UTR (untranslated region), a 2418 bp open reading frame, and a 849 bp 3' UTR. The deduced amino acids of the grass carp SIMP (gcSIMP) are a highly conserved protein with a STT3 domain and 11 transmembrane regions. The gcSIMP spans over more than 24,212 bp in length, containing 16 exons and 15 introns. Most encoding exons, except the first and the 15th, have the same length as those in human and mouse. The gcSIMP promoter contains many putative transcription factor binding sites, such as Oct-1, GCN4, YY1, Sp1, Palpha, TBP, GATA-1, C/EBP beta, and five C/EBP alpha binding sites. The mRNA expression of gcSIMP in different organs was examined by real-time PCR. The gcSIMP was distributed in all the organs examined, with the highest level in brain, followed by the level in the heart, liver, gill, trunk kidney, muscle, head kidney, thymus, and the lowest level in spleen. Furthermore, the recombinant gcSIMP has been constructed successfully and expressed in Escherichia coli by using pQE-40 vector, and the polyclonal antibody for rabbit has been successfully obtained, which was verified to be specific. Identification of gcSIMP will help to explore the function in fish innate immunity. (c) 2007 Elsevier Ltd. All rights reserved.
学科主题	Fisheries; Immunology; Marine & Freshwater Biology; Veterinary Sciences
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
研究领域[WOS]	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
关键词[WOS]	MINOR HISTOCOMPATIBILITY ANTIGENS ; T-LYMPHOCYTE RESPONSES ; N-LINKED GLYCOSYLATION ; CLASS-I ; OLIGOSACCHARYLTRANSFERASE COMPLEX ; YEAST OLIGOSACCHARYLTRANSFERASE ; STT3 PROTEIN ; GENE ; C/EBP ; IDENTIFICATION
收录类别	SCI
语种	英语
WOS记录号	WOS:000257007000004
公开日期	2010-10-13
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/152342/8098]
专题	水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文
作者单位	1.Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China 2.Chinese Acad Sci, Inst Hydrobiol, Lab Fish Dis, Wuhan 430072, Hubei Province, Peoples R China 3.Huazhong Agr Univ, Coll Anim Sci & Vet Med, Wuhan 430072, Peoples R China
推荐引用方式 GB/T 7714	Xu, Z. Y.,Nie, P.,Chang, M. X.,et al. Cloning, characterization and expression analysis of SIMP (source of immunodominant MHC-associated peptides) in grass carp Ctenopharyngodon idella[J]. FISH & SHELLFISH IMMUNOLOGY,2008,24(6):701-714.
APA	Xu, Z. Y.,Nie, P.,Chang, M. X.,&Sun, B. J..(2008).Cloning, characterization and expression analysis of SIMP (source of immunodominant MHC-associated peptides) in grass carp Ctenopharyngodon idella.FISH & SHELLFISH IMMUNOLOGY,24(6),701-714.
MLA	Xu, Z. Y.,et al."Cloning, characterization and expression analysis of SIMP (source of immunodominant MHC-associated peptides) in grass carp Ctenopharyngodon idella".FISH & SHELLFISH IMMUNOLOGY 24.6(2008):701-714.