ZHANG Qiong-Yu, FAN Si-Gang, LUO Chen. SEQUENCE CLON ING AND EXPRESSION ANALYSIS OF RECOMBINATION ACT IVAT ING GENE 1 AND 2( rag1 AND rag2) IN GRASS CARP, CTENOPHARYNGODON IDELLUS[J]. ACTA HYDROBIOLOGICA SINICA, 2009, 33(5): 795-803.
Citation: ZHANG Qiong-Yu, FAN Si-Gang, LUO Chen. SEQUENCE CLON ING AND EXPRESSION ANALYSIS OF RECOMBINATION ACT IVAT ING GENE 1 AND 2( rag1 AND rag2) IN GRASS CARP, CTENOPHARYNGODON IDELLUS[J]. ACTA HYDROBIOLOGICA SINICA, 2009, 33(5): 795-803.

SEQUENCE CLON ING AND EXPRESSION ANALYSIS OF RECOMBINATION ACT IVAT ING GENE 1 AND 2( rag1 AND rag2) IN GRASS CARP, CTENOPHARYNGODON IDELLUS

  • Received Date: December 03, 2008
  • Rev Recd Date: June 08, 2009
  • Published Date: September 24, 2009
  • It is well known that the infectious diseases caused by the virus and bacterium frequently occur in the one-yearand two-year old grass carp but rarely in the three-year old grass carp,suggesting that perfect mi mune system might be establishedin three-year o ld grass carp. Bymediating V( D) J recom bination of the lymphocyte surface receptor gene exclusively,recom bination act ivat ing genes (RAGs) are the key genes play ing p ivotal role in regulating the developm ent of specific mi m un ity system in vertebrate. To understand the developm ental genetics of mi mune system in grass carp,w e hascloned grass carp rag1 and rag2 genesw ith polym erase chain reaction ( PCR) m ethod and then exam ined their expressionin various developm ental stages with method of reverse transcription-po lym erase cha in reaction ( RT-PCR). The totallength of the genomic sequence of the grass carp rag1 gene from the initiation codon to the stop codon is 4188bp,whichcomposed of three exons and two introns. T he length of entire open read ing frame (ORF) is 3192bp,encoding a prote inconta ining 1063 am ino ac ids. The grass carp rag2 gene has no intron and the length of open reading fram e is 1593bp encodinga prote in containing 530 amino acids. A lignm ent of RAGs am inoacids among various vertebrate species has shownthat the putative sequences in predictive function reg ions of grass carp shared high ident ity with other vertebrates. The expression of rag1 in grass carp embryos has been detected at 4 days post-fertilization (dpf),and higher expression level hasbeen detectedm ainta ining during the subsequent larva stage. rag1 expression has been detected in head kidney of one-yearold grass carp butnot in the three-year old grass carp. These results suggest that theV(D) J recom bination of the mimunoglobulin(Ig) and T-cell receptor (TCR) genes and the concom itant differentiation of lymphocyte start at hatching stageand arema intained in larva and one-year o ld grass carp; but in three-year old grass carp,the activation ofV(D) J recombination is greatly reduced meaning the libraries of diversified Ig and TCR and relat ive perfect mimune system are establishedat this stage.
  • [1]
    ?? N iD S, W ang J G. B iology and D iseases of Grass C arp [M]. eijing: A cadem ic Press. 1999, 38?? 41, 329?? 330 [倪达书,建国. 草鱼生物学与疾病. 北京: 科学出版社. 1999, 38??1, 329?? 330]

    ?? N iD S, W ang J G. B iology and D iseases of Grass C arp [M]. eijing: A cadem ic Press. 1999, 38?? 41, 329?? 330 [倪达书,建国. 草鱼生物学与疾病. 北京: 科学出版社. 1999, 38??1, 329?? 330]
    [2]
    ?? Zhang Q Y, X ie J Y, Zhao R R, et al. Prelim inary proteom ic ifference analysis on th e k idney of grass carp, C tenopharyngod ondellus, in various developm en tal s tages [J]. Ac taH yd robiologica inica, 2006, 30( 4 ): 425?? 431 [张琼宇, 谢锦云, 赵如榕,. 不同发育阶段草鱼肾脏蛋白质组差异的初步分析. 水生物学报, 2006, 30( 4 ): 425?? 431]

    ?? Zhang Q Y, X ie J Y, Zhao R R, et al. Prelim inary proteom ic ifference analysis on th e k idney of grass carp, C tenopharyngod ondellus, in various developm en tal s tages [J]. Ac taH yd robiologica inica, 2006, 30( 4 ): 425?? 431 [张琼宇, 谢锦云, 赵如榕,. 不同发育阶段草鱼肾脏蛋白质组差异的初步分析. 水生物学报, 2006, 30( 4 ): 425?? 431]
    [3]
    ?? Van G D, M cb lane J F, Ram sd en D A, et al. In it iation of VD ) J recomb in at ion in a cel-l free system [J]. C ell, 1995, 81:25?? 934
    [4]
    ?? M ark et E, Papavas iliou F N. V( D) J recomb in at ion and the evoluion of the adap tive imm une system [J]. PL oS B iology, 2003,( 1) : e16
    [5]
    ?? L itm an G W, And erson M K, Rast J P. E volut ion of an t igen ind ing recep tors [J]. Annua lR eview of Imm unology, 1999, 17 1) : 109?? 147
    [6]
    ?? B rand tV L, RothD B. A recom b inase d iversif ied: new functionsf the RAG p rote ins [J]. Current Op in ion in Imm unology,002, 14( 2) : 224?? 229
    [7]
    ?? Groth J G, Barrow clough G F. Basal d ivergences in b irds and thehylogenetic ut ility of the nuclear RAG-1 gen e [J]. M olecu larhylog ene tic s and E volu tion, 1999, 12( 2 ): 115?? 123
    [8]
    ?? Lovejoy N R, Co llette B B. Phylogenetic relationsh ip s of n ew orld needlefishes ( Teleos te:i B elon idae) and the b iogeographyf trans it ion s b etw een m arin e and freshw ater hab itats [J]. C opea, 2001, 2001( 2) : 324?? 338
    [9]
    ?? Su llivan J P, Lundb erg JG, H ardm anM. A phylogenetic analysisf the m ajor group s of catf ish es ( Teleoste:i S ilu riform es ) sing rag1 and rag2 nu clear gen e sequen ces [J]. M olecu lar P hylogenetcs and E volu tion, 2006, 41 ( 3) : 636?? 662
    [10]
    ?? Wang X, L i J, H e S. Mo lecu lar evidence for the monophyly ofastA sian groups of Cyp rin idae ( Teleoste:i Cyprin iform es ) derivedrom the nuclear recom b ination activating gen e 2 sequ ences[J]. M olecular Phylogene tic s and E volu tion, 2007, 42 ( 1 ) :57?? 170
    [11]
    ?? H ansen J D, Kaattari S L. The recom b ination act ivat ing gene 1RAG1 ) of rainbow trou t ( Oncorhynchu s mykiss) : clon ing, expess ion, and phylogenetic analysis [J]. Immun og enetics, 1995,2( 3) : 188?? 195
    [12]
    ?? H ansen J, Kaat tari S. Th e recomb in at ion act ivat ing gene 2RAG2 ) of the rainbow trou tOncorhynchu s mykiss [J]. Immunogenetics,996, 44 ( 3) : 203?? 211
    [13]
    ?? W illett C E, Cherry J J, S teiner L A. Characterizat ion and expess ion of the recom b ination act ivat ing gen es ( rag 1 and rag 2) ofebraf ish [J]. Immun og en etics, 1997, 45( 6) : 394?? 404
    [14]
    ?? W illet t C E, Zapata A G, H opk in sN, e t a l. Exp ress ion of zebafish RAG genes during early developm en t iden tif ies the thym us[J]. Developm en ta lB iology, 1997, 182 ( 2) : 331?? 341
    [15]
    ?? Peixoto B R, M ikawa Y, Brenner S. Ch aracterizat ion of theecom binase act ivat ing gene-1 and 2 locus in the Japan ese pu fferfish,ugu rubripes [J]. G ene, 2000, 246( 1?? 2) : 275?? 283
    [16]
    ?? Gong Y W, Zhang Y, H uang Q, et al. C lon ing and id ent if icat ion??802 水?? ?? 生?? ?? 生?? ?? 物?? ?? 学?? ?? 报 33卷f the rag1 gene in channel catfish [J]. Acta Acad em iaeM ed icinaeMl itaris Tertiae, 2004, 26 ( 8 ): 700?? 703 [公衍文, 张, 黄庆, 等. Channel catf ish重组激活基因克隆与鉴定. 第军医大学学报, 2004, 26( 8) : 700?? 703]

    ?? Gong Y W, Zhang Y, H uang Q, et al. C lon ing and id ent if icat ion??802 水?? ?? 生?? ?? 生?? ?? 物?? ?? 学?? ?? 报 33卷f the rag1 gene in channel catfish [J]. Acta Acad em iaeM ed icinaeMl itaris Tertiae, 2004, 26 ( 8 ): 700?? 703 [公衍文, 张, 黄庆, 等. Channel catf ish重组激活基因克隆与鉴定. 第军医大学学报, 2004, 26( 8) : 700?? 703]
    [17]
    ?? H u ttenhuisH B, Hu ising M O, VanM T, et a l. Rag expressionden tif ies B and T cel l lym phopoietic t issues during the developmnt of comm on carp ( Cyprinus carpio) [J]. D eve lopm en ta l&omparative Immunology, 2005, 29 ( 12) : 1033?? 1047
    [18]
    ?? Fan S G, Zhang Q Y, Luo C. Sequ ence clon ing and expressionnalys is of R ag gen es in goldf ish, carass ius auratu s [J]. A ctayd robiolog ica S in ica, 2009, 33 ( 4) : 603?? 612 [范嗣刚, 张琼, 罗琛. 鲫鱼Rag基因的克隆及表达分析. 水生生物学报,009, 33( 4 ): 603?? 612]

    ?? Fan S G, Zhang Q Y, Luo C. Sequ ence clon ing and expressionnalys is of R ag gen es in goldf ish, carass ius auratu s [J]. A ctayd robiolog ica S in ica, 2009, 33 ( 4) : 603?? 612 [范嗣刚, 张琼, 罗琛. 鲫鱼Rag基因的克隆及表达分析. 水生生物学报,009, 33( 4 ): 603?? 612]
    [19]
    ?? Ch en JH, H uang M M, Zh eng K, e t al. RAPD analys is on genomc DNA of two art ificial gynogen et ic groups of grass carp, tenopharyngod on id ellus [J]. A cta H yd robiologica S in ica,004, 28( 5 ): 471?? 477 [陈金辉, 黄明敏, 郑康, 等. 两个同的人工雌核发育草鱼群体基因组DNA 的RAPD 分析.生生物学报, 2004, 28( 5) : 471?? 477]

    ?? Ch en JH, H uang M M, Zh eng K, e t al. RAPD analys is on genomc DNA of two art ificial gynogen et ic groups of grass carp, tenopharyngod on id ellus [J]. A cta H yd robiologica S in ica,004, 28( 5 ): 471?? 477 [陈金辉, 黄明敏, 郑康, 等. 两个同的人工雌核发育草鱼群体基因组DNA 的RAPD 分析.生生物学报, 2004, 28( 5) : 471?? 477]
    [20]
    ?? Sadofsky M J, H essw JE, M cb lane J F, e ta l. Expression and V D) J recomb in at ion activ ity of m u tated RAG-1 protein s [J].u cl. A cid s R es., 1993, 21( 24 ) : 5644?? 5650
    [21]
    ?? M cm ahan C J, Dif ilippan ton ioM J, Rao N, e t al. A basicm ot ifn th eN-term inal reg ion ofRAG1 enhan cesV ( D) J recom b inationct ivity [J]. M ol. C ell. B iol., 1997, 17( 8) : 4544?? 4552
    [22]
    ?? A id in is V, D iasD C, Gom ez C A, e t a l. Def in it ion ofm in im alom ains of interact ion w ith in the recom b inat ion-act ivat ing gen es 1nd 2 recomb in ase comp lex [J]. J Immunol, 2000, 164 ( 11 ):826?? 5832
    [23]
    ?? Sadofsky M J, H esse J E, G ellertM. Defin it ion of a core regionfRAG-2 that is functional inV ( D) J recomb inat ion [J]. Nu cl. cid s R es., 1994, 22( 10) : 1805?? 1809
    [24]
    ?? C uom o C A, O ett ingerM A. An alysis of reg ion s ofRAG-2 importan for V (D ) J recom b inat ion [J]. Nuc l. Acids R es., 1994, 22 10 ): 1810?? 1814
    [25]
    ?? B iga j J, Du lak J, Plytycz B. Lym phoid organ s ofG asterosteus acuea tus [J]. Journa l of F ish B iology, 1987, 31 ( supplem ent) : 233?? 234
    [26]
    ?? Notarangelo L D, V illa A, S chw arzK. RAG and RAG defects[J]. Cu rren tOpinion in Imm unology, 1999, 11( 4 ): 435?? 442
    [27]
    ?? Noordz ij JG, VerkaikN S, H artw ig N G, et al. N-term in al truncatedum an RAG1 protein s can d irectT-cell receptor but not immnoglobu lin gen e rearrangem ents [J]. B lood, 2000, 96 ( 1 ) :03?? 209
    [28]
    ?? S teen S B, H an J, Mundy C, e t al. Roles of the ?? d ispensab le??ortions ofRAG-1 and RAG-2 in V ( D ) J recomb in at ion [J]. ol. Cel l. B iol., 1999, 19 ( 4) : 3010?? 3017
    [29]
    ?? Cortes P, Ye Z S, B alt im ore D. RAG-1 in teractsw ith the repea-td am ino acid m otif of the hum an homo logu e of th e yeast proteinRP1 [J]. P roc Na tl A cad S ci USA, 1994, 91 ( 16 ) :633?? 7637
    [30]
    ?? S chatzDg, Oett ingerM a B D. The V( D) J recomb inat ion act ivatingen e, RAG-1 [J]. C ell, 1989, 59( 6) : 1035?? 1048
    [31]
    ?? RodgersK K, Bu Z, Flem ing K G, et al. A Zin c-b inding dom ainnvolved in the d im erizat ion of RAG1 [J]. Journa l of M olecu lar iology, 1996, 260( 1 ) : 70?? 84
    [32]
    ?? K irch S A, Rathbun G A, Oett ingerM A. Dual role of RAG2 in ( D) J recomb in at ion: catalysis and regulation of ordered Ig genessemb ly [J]. EMBO Jou rnal, 1998, 17( 16) : 4881?? 4886
    [33]
    ?? Barbara C A. A short p ept ide at the C term inu s is respon sib le forh e nu clear local ization of RAG2 [J]. Eu ropean Jou rnal of Immunology,002, 32 ( 7) : 2068?? 2073
    [34]
    ?? Ross A E, Vu icaM, D es iderio S. Overlapp ing signals for protein egradation and nuclear localizat ion define a ro le for in trins icAG-2 nuclear up take in d ivid ing cells [J]. M ol. Ce ll. B iol.,003, 23( 15) : 5308?? 5319
    [35]
    ?? SadofskyM J. The RAG protein s inV ( D) J recomb inat ion: m oreh an just a nuclease [J]. N ucl. Ac id s R es., 2001, 29 ( 7 ) :399?? 1409
    [36]
    ?? Yurchenko V, Xue Z, S adofsky M. Th e RAG1 N-term in al domin is anE 3 ub iqu it in l igase [J]. G enes& Developmen t, 2003,7( 5) : 581?? 585
    [37]
    ?? S adofsky M J. R ecom b ination-act ivat ing gen e p roteins: m ore reguat ion, p lease [J]. Immunolog ical R ev iew s, 2004, 200 ( 1 ) :3?? 89

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