Volume 33 Issue 2
Mar.  2012
Turn off MathJax
Article Contents
KONG Cheng-Jiang, HUANG Zuo-An, CHEN Jiong, SHI Yu-Hong, LU Xin-Jiang. Molecular cloning, sequence analysis and expression of ayu complement component C9 gene. Zoological Research, 2012, (2): 151-157. doi: 10.3724/SP.J.1141.2012.02151
Citation: KONG Cheng-Jiang, HUANG Zuo-An, CHEN Jiong, SHI Yu-Hong, LU Xin-Jiang. Molecular cloning, sequence analysis and expression of ayu complement component C9 gene. Zoological Research, 2012, (2): 151-157. doi: 10.3724/SP.J.1141.2012.02151

Molecular cloning, sequence analysis and expression of ayu complement component C9 gene

doi: 10.3724/SP.J.1141.2012.02151
  • Received Date: 2011-11-21
  • Rev Recd Date: 2011-12-26
  • Publish Date: 2012-04-22
  • C9, a component of the membrane attack complex, participates in the final stage of the complement cascade which lyses foreign organisms by disrupting the integrity of their cell membranes. In the present study, a full-length ayu C9 (aC9) cDNA was cloned which contains 2,125 nucleotides and encodes a protein of 592 amino acids.A signal peptide was deposited in the N-terminal 22 residues. The deduced amino acid sequence of aC9 showed 56.8% identity to the C9 of rainbow trout, and 40.9% to 53.8% identity to the C9 of other teleosts. RT-PCR analysis demonstrated that the mRNA of aC9 was expressed in the liver, spleen, intestine, gill and muscle of healthy ayu fish with the highest level in the liver. Quantitative RT-PCR analysis showed that aC9 transcripts were significantly up-regulated in the liver at 4 h post Listonella anguillarum infection, peaked at 16 h post injection. Western blotting analysis revealed that serum aC9 significantly increased in Listonella anguillarum infected ayu fish. Our results suggested that aC9 may play an important role in fish immune response of anti-bacteria.
  • loading
  • [1]
    Biesecker G, Gerard C, Hugli TE. 1982. An amphiphilic structure of the ninth component of human complement. Evidence from analysis of fragments produced by alpha-thrombin[J]. J Biol Chem, 257(5): 2584-2590.
    Bunkenborg J, Pilch BJ, Podtelejnikov AV, Wi niewski JR. 2004. Screening for N-glycosylated proteins by liquid chromatography mass spectrometry[J]. Proteomics, 4(2): 454-465.
    Chen J, Shi YH, Li MY, Ding WC, Niu H. 2008. Molecular cloning of liver angiotensinogen gene in ayu (Plecoglossus altivelis) and mRNA expression changes upon Aeromonas hydrophila infection[J]. Fish Shellfish Immunol, 24(5): 659-662.
    Chondrou MP, Londou AV, Zarkadis IK. 2006. Expression and phylogenetic analysis of the ninth complement component (C9) in rainbow trout[J]. Fish Shellfish Immunol, 21(5): 572-576.
    Dunkelberger JR, Song WC. 2010. Complement and its role in innate and adaptive immune responses[J]. Cell Res, 20(1): 34-50.
    Encinas P, Rodriguez-Milla MA, Novoa B, Estepa A, Figueras A, Coll J. 2010. Zebrafish fin immune responses during high mortality infections with viral haemorrhagic septicemia rhabdovirus. A proteomic and transcriptomic approach[J]. BMC Genomics, 11: 518.
    Endo Y, Matsushita M, Fujita T. 2011. The role of ficolins in the lectin pathway of innate immunity[J]. Int J Biochem Cell Biol, 43(5): 705-712.
    Fosbrink M, Niculescu F, Rus H. 2005. The role of c5b-9 terminal complement complex in activation of the cell cycle and transcription[J]. Immunol Res, 31(1): 37-46.
    Gerwick L, Steinhauer R, Lapatra S, Sandell T, Ortuno J, Hajiseyedjavadi N, Bayne CJ. 2002. The acute phase response of rainbow trout (Oncorhynchus mykiss) plasma proteins to viral, bacterial and fungal inflammatory agents[J]. Fish Shellfish Immunol, 12(3): 229-242.
    Huang ZA, Chen J, Lu XJ, Shi YH, Li MY. 2011. Alteration on the expression of ayu coagulation factor X gene upon Listonella anguillarum infection[J]. Zool Res, 32(5): 492-498. [黄左安, 陈炯, 陆新江, 史雨红, 李明云. 2011. 香鱼凝血因子 X 基因表达与鳗利斯顿氏菌感染的相关性[J]. 动物学研究, 32(5): 492-498.]
    Katagiri T, Hirono I, Aoki T. 1999. Molecular analysis of complement component C8beta and C9 cDNAs of Japanese flounder, Paralichthys olivaceus[J]. Immunogenetics, 50(1-2): 43-48.
    Li CH, Chen J, Shi YH, Li MY. 2009. Characterization of Listonella anguillarum as the aetiological agent of vibriosis occurred in cultured ayu (Plecoglossus altivelis) in Ninghai country, China[J]. Acta Micriobiol Sin, 2009, 49(7): 931-937. [李长红, 陈炯, 史雨红, 李明云. 2009. 宁海地区香鱼弧菌病病原菌鉴定[J]. 微生物学报, 49(7): 931-937.]
    Li L, Chang MX, Nie P. 2007. Molecular cloning, promoter analysis and induced expression of the complement component C9 gene in the grass carp Ctenopharyngodon idella[J]. Vet Immunol Immunopathol, 118(3-4): 270-282.
    Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J]. Methods, 25(4): 402-408.
    Muller-Eberhard HJ. 1986. The membrane attack complex of complement[J]. Annu Rev Immunol, 4(1): 503-528.
    Qu HC, Ricklin D, Lambris JD. 2009. Recent developments in low molecular weight complement inhibitors[J]. Mol Immunol, 47(2-3): 185-195.
    Rutkowski MJ, Sughrue ME, Kane AJ, Ahn BJ, Fang SN, Parsa AT. 2010. The complement cascade as a mediator of tissue growth and regeneration[J]. Inflamm Res, 59(11): 897-905.
    Sarmar JV, Ward PA. 2011. The complement system[J]. Cell Tissue Res, 343(1): 227-235.
    Shi YH, Chen J, Li CH, Li MY. 2010. Molecular cloning of liver Wap65 cDNA in ayu (Plecoglossus altivelis) and mRNA expression changes following Listonella anguillarum infection[J]. Mol Biol Rep, 37(3): 1523-1529.
    Stanley KK, Herz J. 1987. Topological mapping of complement component C9 by recombinant DNA techniques suggests a novel mechanism for its insertion into target membranes[J]. EMBO J, 6(7): 1951-1957.
    Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Mol Biol Evol, 24(8): 1596-1599.
    Toapanta FR, Ross TM. 2006. Complement-mediated activation of the adaptive immune responses: role of C3d in linking the innate and adaptive immunity[J]. Immunol Res, 36(1-3): 197-210.
    Tomlinson S, Stanley KK, Esser AF. 1993. Domain structure, functional activity, and polymerization of trout complement protein C9[J]. Dev Comp Immunol, 17(1): 67-76.
    Uenobe M, Kohchi C, Yoshioka N, Yuasa A, Inagawa H, Morii K, Nishizawa T, Takahashi Y, Soma GI. 2007. Cloning and characterization of a TNF-like protein of Plecoglossus altivelis (ayu fish)[J]. Mol Immunol, 44(6): 1115-1122.
    Wagner E, Frank MM. 2010. Therapeutic potential of complement modulation[J]. Nat Rev Drug Discov, 9(1): 43-56.
    Witzel-Schlomp K, Rittner C, Schneider PM. 2001. The human complement C9 gene: structural analysis of the 5’ gene region and genetic polymorphism studies[J]. Eur J Immunogenet, 28(5): 515-522.
    Yeo GSH, Elgar G, Sandford R, Breneer S. 1997. Cloning and sequencing of complement component C9 and its linkage to DOC-2 in the pufferfish Fugu rubripes[J]. Gene, 200(1-2): 203-211.
  • 加载中


    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Article Metrics

    Article views (1976) PDF downloads(1424) Cited by()
    Proportional views


    DownLoad:  Full-Size Img  PowerPoint