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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; Nie, P, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China
2008-06-01
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume24Issue:6Pages:701-714
AbstractSIMP (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.; 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.
SubtypeArticle
KeywordSimp Grass Carp Cdna Gene Organization Promoter Organ
Department[Xu, Z. Y.; Nie, P.; Chang, M. X.; Sun, B. J.] Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China; [Xu, Z. Y.; Nie, P.; Chang, M. X.; Sun, B. J.] Chinese Acad Sci, Inst Hydrobiol, Lab Fish Dis, Wuhan 430072, Hubei Province, Peoples R China; [Xu, Z. Y.] Huazhong Agr Univ, Coll Anim Sci & Vet Med, Wuhan 430072, Peoples R China
Subject AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
DOI10.1016/j.fsi.2007.11.019
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS SubjectFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS IDWOS:000257007000004
WOS KeywordMINOR HISTOCOMPATIBILITY ANTIGENS ; T-LYMPHOCYTE RESPONSES ; N-LINKED GLYCOSYLATION ; CLASS-I ; OLIGOSACCHARYLTRANSFERASE COMPLEX ; YEAST OLIGOSACCHARYLTRANSFERASE ; STT3 PROTEIN ; GENE ; C/EBP ; IDENTIFICATION
Citation statistics
Cited Times:5[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8098
Collection期刊论文
Corresponding AuthorNie, P, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China
Affiliation1.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
Recommended Citation
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.,&Nie, P, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China.(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.
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