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Cloning, characterization, and expression analysis of a CC chemokine gene from turbot (Scophthalmus maximus)

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Abstract

The chemokines are a superfamily of chemotactic cytokines playing an important role in leukocyte chemotaxis. Here, a turbot head kidney cDNA library was constructed in which KC70 was identified as a CC chemokine. Unknown 5′ and 3′ parts of the cDNA were amplified by 5′ and 3′ rapid amplification of cDNA ends (RACE). The complete cDNA of KC70 contains a 59-bp 5′ UTR, a 336-bp ORF, and a 152-bp 3′ UTR. Four exons and three introns were identified in KC70. Phylogenetic analysis showed that KC70 was similar to CCL19. In normal turbot KC70 was expressed in all tissues except brain and skin. Infection of turbot with pathogenic bacteria significantly increased expression of KC70 in the liver. Expression of KC70 in head kidney first increased and then decreased after bacterial challenge. No significant change was observed in the spleen after bacterial challenge. During embryonic development, KC70 was highly expressed after the gastrula stage. These results indicated KC70 plays important and multiple roles in turbot immune response.

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Abbreviations

TECs:

Turbot embryonic cell line

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse transcription PCR

ORF:

Open reading frame

UTR:

Untranslated region

References

  • Alexander JB, Ingram GA (1992) Noncellular nonspecific defense mechanisms of fish. Annu Rev Fish Dis 2:249–279

    Article  Google Scholar 

  • Bacon K, Baggiolini M, Broxmeyer H, Horuk R, Lindley I, Mantovani A, Matsushima K, Murphy P, Nomiyama H, Oppenheim J, Rot A, Schall T, Tsang M, Thorpe R, Van Damme J, Wadhwa M, Yoshie O, Zlotnik A, Zoon K (2003) Chemokine/chemokine receptor nomenclature. Cytokine 21:48–49

    Article  CAS  Google Scholar 

  • Bao B, Peatman E, Peng X, Baoprasertkul P, Wang G, Liu Z (2006) Characterization of 23 CC chemokine genes and analysis of their expression in channel catfish (Ictalurus punctatus). Dev Comp Immunol 30:783–796

    Article  CAS  PubMed  Google Scholar 

  • Chen SL, Hong Y, Scherer S, Schartl M (2001) Lack of ultraviolet-light inducibility of the medakafish (Oryzias latipes) tumor suppressor gene p53. Gene 264:197–203

    Article  CAS  PubMed  Google Scholar 

  • Chen SL, Xu MY, Hu SN, Li L (2004) Analysis of immune-relevant genes expressed in red sea bream (Chrysophrys major) spleen. Aquaculture 240:115–130

    Article  CAS  Google Scholar 

  • Chen J, Li C, Yan X, Li Y, Chi Z, Xue X, Wang Y (2005) Studies on biological characteristics of five pathogenic Vibrio anguillarum strains isolated from diseased turbot (Scophthalmus maximus) in Shandong province of China. Chinese High Technol Lett 15(6):92–96

    CAS  Google Scholar 

  • Chen SL, Li W, Meng L, Sha ZX, Wang ZJ, Ren GC (2007) Molecular cloning and expression analysis of a hepcidin antimicrobial peptide gene from turbot (Scophthalmus maximus). Fish Shellfish Immunol 22:172–181

    Article  PubMed  Google Scholar 

  • Dayhoff MO, Schwartz RM, Orcutt BC (1978) A model of evolutionary change. In: Dayhoff MO (ed) Atlas of protein sequence and structure, vol 5, Suppl 3. Washington, DC: National Biomedical Research Foundation, pp 345–358

  • Devries ME, Kelvin AA, Xu L, Ran L, Robinson J, Kelvin DJ (2006) Defining the origins and evolution of the chemokine/chemokine receptor system. J Immunol 176:401–415

    CAS  PubMed  Google Scholar 

  • Du Pasquier L (1982) Antibody diversity in lower vertebrates—why is it so restricted? Nature 296:311–313

    Article  PubMed  Google Scholar 

  • Dixon B, Shum B, Adams EJ, Magor KE, Hedrick RP, Muir DG, Parham P (1998) CK-1, a putative chemokine of rainbow trout (Oncorhynchus mykiss). Immunol Rev 166:341–348

    Article  CAS  PubMed  Google Scholar 

  • Ellis AE (2001) Innate host defence mechanism of fish against viruses and bacteria. Dev Comp Immunol 25:827–839

    Article  CAS  PubMed  Google Scholar 

  • Fujiki K, Shin DH, Nakao M, Yano T (1999) Molecular cloning of carp (Cyprinus carpio) CC chemokine, CXC chemokine receptors, allograft inflammatory factor-1, and natural killer cell enhancing factor by use of suppression subtractive hybridization. Immunogenetics 49:909–914

    Article  CAS  PubMed  Google Scholar 

  • He C, Peatman E, Baoprasertkul P, Kucuktas H, Liu ZJ (2004) Multiple CC chemokines in channel catfish and blue catfish as revealed by analysis of expressed sequence tags. Immunogenetics 56:379–387

    Article  CAS  PubMed  Google Scholar 

  • Kuroda N, Uinuk-ool TS, Sato A, Samonte IE, Figueroa F, Mayer WE, Klein J (2003) Identification of chemokines and a chemokine receptor in cichlid fish, shark, and lamprey. Immunogenetics 54:884–895

    CAS  PubMed  Google Scholar 

  • Kumar S, Tamura K, Nei M (2004) MEGA3.0: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163

    Article  CAS  PubMed  Google Scholar 

  • Laing KJ, Secombes CJ (2004a) Chemokines. Dev Comp Immunol 28:443–460

    Article  CAS  PubMed  Google Scholar 

  • Laing KJ, Secombes CJ (2004b) Trout CC chemokines: comparison of their sequences and expression patterns. Mol Immunol 41:793–808

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Chen SL, Meng L, Zhang YX (2007) Cloning, characterization and expression analysis of a novel CXC chemokine from turbot (Scophthalmus maximus). Fish Shellfish Immunol 23:711–720

    Article  CAS  PubMed  Google Scholar 

  • Magnadottir B (2006) Innate immunity of fish. Fish Shellfish Immunol 20:137–151

    Article  CAS  PubMed  Google Scholar 

  • Nielsen H, Engelbrencht J, Brunak S, Heijne G (1997) Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10:1–6

    Article  CAS  PubMed  Google Scholar 

  • Press CM, Evensen (1999) The morphology of the immune system in teleost fishes. Fish Shellfish Immunol 9:309–318

  • Peatman E, Bao BL, Baoprasertkul P, Liu ZJ (2005) In silico identification and expression analysis of 12 novel CC chemokines in catfish. Immunogenetics 57:409–419

    Article  CAS  PubMed  Google Scholar 

  • Peatman E, Bao B, Peng X, Baoprasertkul P, Brady Y, Liu ZJ (2006) Catfish CC chemokines: genomic clustering, duplications, and expression after bacterial infection with Edwardsiella ictaluri. Mol Gen Genom 275:297–309

    Article  CAS  Google Scholar 

  • Peatman E, Liu ZJ (2006) CC chemokines in zebrafish: evidence for extensive intrachromosomal gene duplications. Genomics 88:381–385

    Article  CAS  PubMed  Google Scholar 

  • Peatman E, Liu ZJ (2007) Evolution of CC chemokines in teleost fish: a case study in gene duplication and implications for immune diversity. Immunogenetics 59:613–623

    Article  CAS  PubMed  Google Scholar 

  • Rutch K, Tsuyoshi O , Ikuo H, Takashi A (2004) Identification of a novel Japanese flounder (Paralichthys olivaceus) CC chemokine gene and an analysis of its function. Immunogenetics 55:763–769

    Article  Google Scholar 

  • Waterston RH, Lindblad-Toh K, Birney E, Rogers J, Abril JF, Agarwal P, Agarwala R, Ainscough R, Alexandersson M, An P, Birren B, Barlow K, Baertsch R, Beck S, Abril JF, Ainscough R, Antonarakis SE, Bloom T, Bailey J, Attwood J (2002) Initial sequencing and comparative analysis of the mouse genome. Nature 420:520–562

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from National Major Basic Research Program (973) of China (2004CB117403), 863 Hightech project (2006AA10A402), National Nature Science Foundation of China (40376047, 30671623), and Taishan Scholar Project of Shandong Province.

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Correspondence to S. L. Chen.

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Chen, S.L., Liu, Y., Dong, X.L. et al. Cloning, characterization, and expression analysis of a CC chemokine gene from turbot (Scophthalmus maximus). Fish Physiol Biochem 36, 147–155 (2010). https://doi.org/10.1007/s10695-008-9218-1

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  • DOI: https://doi.org/10.1007/s10695-008-9218-1

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