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Structure, organization and expression of common carp (Cyprinus carpio L.) SLP-76 gene
Huang, Rong1,2; Sun, Xiao-Feng1,2; Hu, Wei1; Wang, Ya-Ping1; Guo, Qiong-Lin1; Guo, QL, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2008-05-01
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume24Issue:5Pages:530-541
AbstractSLP-76 is an important member of the SLP-76 family of adapters, and it plays a key role in TCR signaling and T cell function. Partial cDNA sequence of SLP-76 of common carp (Cyprinus carpio L.) was isolated from thymus cDNA Library by the method of suppression subtractive hybridization (SSH). Subsequently, the full length cDNA of carp SLP-76 was obtained by means of 3' RACE and 5' RACE, respectively. The full Length cDNA of carp SLP-76 was 2007 bp, consisting of a T-terminal untranslated region (UTR) of 285 bp, a T-terminal. UTR of 240 bp, and an open reading frame of 1482 bp. Sequence comparison showed that the deduced amino acid sequence of carp SLP-76 had an overall similarity of 34-73% to that of other species homotogues, and it was composed of an NH2-terminal domain, a central proline-rich domain, and a C-terminal SH2 domain. Amino acid sequence analysis indicated the existence of a Gads binding site R-X-X-K, a 10-aa-long sequence which binds to the SH3 domain of LCK in vitro, and three conserved tyrosine-containing sequence in the NH2-terminal domain. Then we used PCR to obtain a genomic DNA which covers the entire coding region of carp SLP-76. In the 9.2 k-long genomic sequence, twenty one exons and twenty introns were identified. RT-PCR results showed that carp SLP-76 was expressed predominantly in hematopoietic tissues, and was upregulated in thymus tissue of four-month carp compared to one-year old carp. RT-PCR and virtual northern hybridization results showed that carp SLP-76 was also upregulated in thymus tissue of GH transgenic carp at the age of four-months. These results suggest that the expression level of SLP-76 gene may be related to thymocyte development in teleosts. (c) 2007 Published by Elsevier Ltd.; SLP-76 is an important member of the SLP-76 family of adapters, and it plays a key role in TCR signaling and T cell function. Partial cDNA sequence of SLP-76 of common carp (Cyprinus carpio L.) was isolated from thymus cDNA Library by the method of suppression subtractive hybridization (SSH). Subsequently, the full length cDNA of carp SLP-76 was obtained by means of 3' RACE and 5' RACE, respectively. The full Length cDNA of carp SLP-76 was 2007 bp, consisting of a T-terminal untranslated region (UTR) of 285 bp, a T-terminal. UTR of 240 bp, and an open reading frame of 1482 bp. Sequence comparison showed that the deduced amino acid sequence of carp SLP-76 had an overall similarity of 34-73% to that of other species homotogues, and it was composed of an NH2-terminal domain, a central proline-rich domain, and a C-terminal SH2 domain. Amino acid sequence analysis indicated the existence of a Gads binding site R-X-X-K, a 10-aa-long sequence which binds to the SH3 domain of LCK in vitro, and three conserved tyrosine-containing sequence in the NH2-terminal domain. Then we used PCR to obtain a genomic DNA which covers the entire coding region of carp SLP-76. In the 9.2 k-long genomic sequence, twenty one exons and twenty introns were identified. RT-PCR results showed that carp SLP-76 was expressed predominantly in hematopoietic tissues, and was upregulated in thymus tissue of four-month carp compared to one-year old carp. RT-PCR and virtual northern hybridization results showed that carp SLP-76 was also upregulated in thymus tissue of GH transgenic carp at the age of four-months. These results suggest that the expression level of SLP-76 gene may be related to thymocyte development in teleosts. (c) 2007 Published by Elsevier Ltd.
SubtypeArticle
KeywordSlp-76 Race Molecular Cloning Common Carp
Department[Huang, Rong; Sun, Xiao-Feng; Hu, Wei; Wang, Ya-Ping; Guo, Qiong-Lin] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; [Huang, Rong; Sun, Xiao-Feng] Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Subject AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
DOI10.1016/j.fsi.2007.10.011
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:000255675700006
WOS KeywordT-CELL DEVELOPMENT ; TYROSINE-PHOSPHORYLATED SLP-76 ; ADAPTER PROTEIN SLP-76 ; LINKER PROTEIN ; BINDING-SITE ; DIFFERENTIAL REQUIREMENT ; MOLECULAR-CLONING ; ANTIGEN RECEPTOR ; IGE RECEPTOR ; RXXK MOTIF
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8148
Collection期刊论文
Corresponding AuthorGuo, QL, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Affiliation1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Recommended Citation
GB/T 7714
Huang, Rong,Sun, Xiao-Feng,Hu, Wei,et al. Structure, organization and expression of common carp (Cyprinus carpio L.) SLP-76 gene[J]. FISH & SHELLFISH IMMUNOLOGY,2008,24(5):530-541.
APA Huang, Rong,Sun, Xiao-Feng,Hu, Wei,Wang, Ya-Ping,Guo, Qiong-Lin,&Guo, QL, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(2008).Structure, organization and expression of common carp (Cyprinus carpio L.) SLP-76 gene.FISH & SHELLFISH IMMUNOLOGY,24(5),530-541.
MLA Huang, Rong,et al."Structure, organization and expression of common carp (Cyprinus carpio L.) SLP-76 gene".FISH & SHELLFISH IMMUNOLOGY 24.5(2008):530-541.
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