鱼类IL-1β和IL-1R1-like基因的分子克隆和鉴定
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摘要
白细胞介素-1(Interleukin-1,IL-1)是在局部和全身炎症反应中起关键作用的细胞因子,包括IL-1α和IL-1β(又分别被命名为IL-1F1和IL-1F2)。目前,IL-1家族在哺乳动物中共发现了11个因子,包括IL-1F1-IL-1F11,但是在鱼类或其他低等脊椎动物中,除IL-1β、IL-18和IL-1RA之外的家族成员尚未被发现。IL-1的生物学效应通过结合其受体,即IL-1受体(IL-1 Receptor)后的信号转导来实现。IL-1受体家族包括Ⅰ型受体(IL-1R1)、Ⅱ型受体(IL-1R2)和IL-1受体辅助蛋白(IL-IRAcP)。
本文利用生物信息学技术,在河豚鱼(Tetraodon nigroviridis)中,成功地预测并克隆了IL-1β基因和IL-1R1样基因(Tetraodon IL-1R1-Like),并且利用DNA重组技术构建了TnIL-1β原核表达载体,转化表达菌株BL21,经IPTG诱导,成功表达并纯化到TnIL-1β蛋白,制备了TnIL-1β兔抗血清。TnIL-1β含有4个外显子,3个内含子,开放阅读框(ORF)长度为774bp,编码257个氨基酸。启动子区域含有TATA Box、NF-KB、CEBP、AP1、CCAAT等转录因子结合位点等,3'UTR含有5个ATTTA instability motif。将TnIL-1β的ORF亚克隆到原核表达载体pET41a上,成功构建了重组表达载体TnIL-1β-pET41a。用IPTG诱导使表达菌株BL21大量表达TnIL-1β蛋白,Ni-NTA纯化后能够得到分子量为60.1Kd的纯化条带。TnIL-1R1-Like基因共有11个外显子,ORF长度为1902bp,编码633个氨基酸,含有3个Ig Domain,1个跨膜区,1个TIR Domain。
综上,本研究为后续开展鱼类IL-1和IL-1R的功能和相互作用关系研究奠定了基础,对研究鱼类免疫系统功能和基因进化具有重要的意义。
Interleukin-1 plays an important role in the local and systemic inflammatory response;including IL-1αand IL-1β(named as IL-1F1 and IL-1F2,respectively).At present,the Interleukin-1 family arised in vertebrates consists of 11 known genes encoding ligands(IL-1F1 to IL-1F11).But to date,no orthologue to IL-1βis found in fish or other low vertebrates.IL-1 functions by binding their receptors,IL-1 Receptor,to mediate signal transduction to realize their biological effects.IL-1 Receptor family contains typeⅠreceptors(IL-1R1),typeⅡreceptors(IL-1R2),and the IL-1 receptor accessory protein(IL-1RAP).
In this study,we developed bioinformatics methods based on genomic databases, successfully cloned IL-1βgene and IL-1R-like gene in Tetraodon,and obtained recombinant TNIL-1βprotein and rabbit anti-IL-1βserum.TNIL-1βcDNA contains 4 exons,3 introns and a 774bp open reading frame(ORF).The peptide deduced from ORF is 257aa in size.TNIL-1βpromoter comprises several transcription binding sites, such as TATA-box,NF-KB,CEBP,AP1,CCAAT,and so on,3'UTR contains 5 ATTTA instability motif.A recombined plasmid TNIL-1β-pET41a was obtained by sub-cloning the ORF of TNIL-1βinto prokaryotic expression plasmid pET41a and sequenced.Target 60.1Kd protein was expressed after IPTG induce and purified by Ni-NTA.TNIL-1R-like gene contains 11 exons,and a 1902bp ORF,coding 633aa, which has 3 Ig Domain,1 transmembrane domain(TMD),and 1 TIR Domain.
All the results lay foundation in function studies of fish IL-1βand IL-1R,and is important for fish immune system research and evolution of immunology genes.
本文利用生物信息学技术,在河豚鱼(Tetraodon nigroviridis)中,成功地预测并克隆了IL-1β基因和IL-1R1样基因(Tetraodon IL-1R1-Like),并且利用DNA重组技术构建了TnIL-1β原核表达载体,转化表达菌株BL21,经IPTG诱导,成功表达并纯化到TnIL-1β蛋白,制备了TnIL-1β兔抗血清。TnIL-1β含有4个外显子,3个内含子,开放阅读框(ORF)长度为774bp,编码257个氨基酸。启动子区域含有TATA Box、NF-KB、CEBP、AP1、CCAAT等转录因子结合位点等,3'UTR含有5个ATTTA instability motif。将TnIL-1β的ORF亚克隆到原核表达载体pET41a上,成功构建了重组表达载体TnIL-1β-pET41a。用IPTG诱导使表达菌株BL21大量表达TnIL-1β蛋白,Ni-NTA纯化后能够得到分子量为60.1Kd的纯化条带。TnIL-1R1-Like基因共有11个外显子,ORF长度为1902bp,编码633个氨基酸,含有3个Ig Domain,1个跨膜区,1个TIR Domain。
综上,本研究为后续开展鱼类IL-1和IL-1R的功能和相互作用关系研究奠定了基础,对研究鱼类免疫系统功能和基因进化具有重要的意义。
Interleukin-1 plays an important role in the local and systemic inflammatory response;including IL-1αand IL-1β(named as IL-1F1 and IL-1F2,respectively).At present,the Interleukin-1 family arised in vertebrates consists of 11 known genes encoding ligands(IL-1F1 to IL-1F11).But to date,no orthologue to IL-1βis found in fish or other low vertebrates.IL-1 functions by binding their receptors,IL-1 Receptor,to mediate signal transduction to realize their biological effects.IL-1 Receptor family contains typeⅠreceptors(IL-1R1),typeⅡreceptors(IL-1R2),and the IL-1 receptor accessory protein(IL-1RAP).
In this study,we developed bioinformatics methods based on genomic databases, successfully cloned IL-1βgene and IL-1R-like gene in Tetraodon,and obtained recombinant TNIL-1βprotein and rabbit anti-IL-1βserum.TNIL-1βcDNA contains 4 exons,3 introns and a 774bp open reading frame(ORF).The peptide deduced from ORF is 257aa in size.TNIL-1βpromoter comprises several transcription binding sites, such as TATA-box,NF-KB,CEBP,AP1,CCAAT,and so on,3'UTR contains 5 ATTTA instability motif.A recombined plasmid TNIL-1β-pET41a was obtained by sub-cloning the ORF of TNIL-1βinto prokaryotic expression plasmid pET41a and sequenced.Target 60.1Kd protein was expressed after IPTG induce and purified by Ni-NTA.TNIL-1R-like gene contains 11 exons,and a 1902bp ORF,coding 633aa, which has 3 Ig Domain,1 transmembrane domain(TMD),and 1 TIR Domain.
All the results lay foundation in function studies of fish IL-1βand IL-1R,and is important for fish immune system research and evolution of immunology genes.
引文
[1]Dinarello CA(1994)."The interleukin-1 family:10 years of discovery".Faseb J.8(15):1314-25.
[2]Barksby HE,Lea SR,Preshaw PM,Taylor JJ:The expanding family of interleukin-1 cytokines and their role in destructive inflammatory disorders.Clin Exp Immunol 2007,149(2):217-225.
[3]Nicola,N.A.(1994).Guidebook to cytokines and their receptors.Oxford."Sambrook and Tooze.
[4]Roux-Lombard.P.The interleukin-1 family.Eur Cytokine Netw 1998,9(4):565-576.
[5]Morgan MM,Clayton CC,Heinricher MM(2004)."Dissociation of hyperalgesia from fever following intracerebroventricular administration of interleukin-lbeta in the rat".Brain Res.1022(1-2):96-100.
[6]Vigers,G.P.A.,Anderson,L.J.,Caes,P.& Brandhuber,B.J.(1997).Crystal structure of the type-1 interleukin-1 receptor complexed with interleukin-1 a.Nature 386,190-194
[7]Ellsaesser,C.F.& Clem,L.W.(1994).Functionally distinct high and low molecular weight species of channel catfish and mouse IL-1.Cytokine 6,10-20
[8]Verburg van Kemenade,B.M.L.,Weyts,F.A.A.,Debets,R.& Flik,G.(1995).Carp macrophages and neutrophilic granulocytes secrete an interleukin-1-like factor.Developmental and Comparative Immunology 19,59-70.
[9]Beuscher H,Gunther C,Rollinghoff M.IL-1 beta is secreted by activated murine macrophages as biologically inactive precursor.J Immunol 1990;144:2179-83.
[10]Wieczorek M.Various aspects of IL-1 biological activity.Ⅱ.IL-1 beta in diseases and the Central Nervous System.Wiad Parazytol.2009;55(2):115-24.
[11]Iizumi T.,Sato S.,Iiyama T.,Hata R.,Amemiya H.,Tomomasa H.,Yazaki T.,Umeda T.Recombinant human interleukin-1 beta analogue as a regulator of hematopoiesis in patients receiving chemotherapy for urogenital cancers.Cancer 1991;68:1520-1523
[12]Anju S.Moolwaney and Orisa J.Igwe.Regulation of the cyclooxygenase-2system by interleukin-1β through mitogen-activated protein kinase signaling pathways:A comparative study of human neuroglioma and neuroblastoma cells.Brain Res Mol Brain Res,2005,137(1-2):202-212.
[13]Javier Duque,ManuelD.Diaz-Munoz,Manuel Fresno and Miguel A.Iniguez.Up-regulation of cyclooxygenase-2 by interleukin-1β in colon carcinoma cells.Cellular Signal,2006,18(8),1262-1269.
[14]Partha Mukhopadhyay,M.Aktar Ali,Animesh Nandi,Peter Carreon,Hak Choy and Debabrata Saha.The Cyclin-Dependent Kinase 2 Inhibitor Down-regulates Interleukin-1β-Mediated Induction of Cyclooxygenase-2 Expression in Human Lung.Carcinoma Cells Cancer Res 2006;66(3):1758-66.
[15]Yun-Jin Jung,Jennifer S.Isaacs,Sunmin Lee,Jane Trepel,and Len Neckers.IL-1β mediated up-regulation of HIF-1α,via an NFkB/COX-2 pathway identifies HIF-1 as a critical link between inflammation and oncogenesis.The FASEB Journal Express Article 2003,17(14):2115-211.
[16]Gabay,C.,Porter,B.,Fantuzzi,G.& Arend,W.P.(1997).Mouse IL-1receptor antagonist isoforms:complementary DNA cloning and protein expression of intracellular isoform and tissue distribution of secreted and intracellular IL-1 receptor antagonist in vivo.Journal of Immunology 159,5905-5913.
[17]Hughes,A.L.(1994).Evolution of the interleukin-1 gene family in mammals.Journal of Molecular Evolution 39,6-12.
[18]Reddy,S A;Huang J H,Liao W S(Nov.1997)."Phosphatidylinositol 3-kinase in interleukin 1 signaling.Physical interaction with the interleukin 1 receptor and requirement in NFkappaB and AP-1 activation".J.Biol.Chem.(UNITED STATES) 272(46):29167-73.
[19]Burns,K;Clatworthy J,Martin L,Martinon F,Plumpton C,Maschera B, Lewis A,Ray K,Tschopp J,Volpe F(Jun.2000)."Tollip,a new component of the IL-1RI pathway,links IRAK to the IL-1 receptor".Nat.Cell Biol.(ENGLAND) 2(6):346-51.
[20]Muzio,M;Ni J,Feng P,Dixit V M(Nov.1997)."IRAK(Pelle) family member IRAK-2 and MyD88 as proximal mediators of IL-1 signaling".Science (UNITED STATES) 278(5343):1612-5.
[21]Huang,J;Gao X,Li S,Cao Z(Nov.1997)."Recruitment of IRAK to the interleukin 1 receptor complex requires interleukin 1 receptor accessory protein".Proc.Natl.Acad.Sci.U.S.A.(UNITED STATES) 94(24):12829-32.
[22]Copeland NG,Silan CM,Kingsley DM,et al.(1991)."Chromosomal location of murine and human IL-1 receptor genes.".Genomics 9(1):44-50.
[23]Dale M,Nicklin MJ(1999)."Interleukin-1 receptor cluster:gene organization of IL1R2,IL1R1,IL1RL2(IL-1Rrp2),IL1RL1(T1/ST2),and IL18R1(IL-1Rrp) on human chromosome 2q.".Genomics 57(1):177-9.
[24]Sims JE,Acres RB,Grubin CE,et al.(1989)."Cloning the interleukin 1receptor from human T cells.".Proc.Natl.Acad.Sci.U.S.A.86(22):8946-50.
[25]Muzio M,Ni J,Feng P,Dixit VM(1997)."IRAK(Pelle) family member IRAK-2 and MyD88 as proximal mediators of IL-1 signaling.".Science 278(5343):1612-5.
[26]Burns K,Clatworthy J,Martin L,et al.(2000)."Tollip,a new component of the IL-1RI pathway,links IRAK to the IL-1 receptor.".Nat.Cell Biol.2(6):346-51.
[27]Reddy SA,Huang JH,Liao WS(1997).Phosphatidylinositol 3-kinase in interleukin 1 signaling.Physical interaction with the interleukin 1 receptor and requirement in NFkappaB and AP-1 activation
[28]Sizemore N,Leung S,Stark GR(1999)."Activation of phosphatidylinositol 3-kinase in response to interleukin-1 leads to phosphorylation and activation of the NF-kappaB p65/RelA subunit.".Mol.Cell.Biol.19(7):4798-805.
[29]N.Morton.Interleukin-1β activates a short STAT-3 isoform in clonal insulin-secreting cells.FEBS Letters,Volume 442,Issue 1,Pages 57-60
[30]Dinarello CA.Interleukin-1,interleukin-1 receptors and interleukin-1receptor antagonist.Int Rev Immunol 1998,16(5-6):457-499.
[31]C.A.Dinarello.The interleukin-1 family:10 years of discovery.FASEB Journal.Volume 8,Issue 15,1994,pages 1314-25.
[32]Arend WP,Malyak M,Guthridge CJ,Gabay C(1998)."Interleukin-1receptor antagonist:role in biology.".Annu.Rev.Immunol.16:27-55.
[33]Huising MO,Stet RJ,Savelkoul HF,Verburg-van Kemenade BM(2004)."The molecular evolution of the interleukin-1 family of cytokines;IL-18 in teleost fish".Dev.Comp.Immunol.28(5):395-413.
[34]Sims JE,Nicklin MJ,Bazan JF,Barton JL,Busfield SJ,Ford JE,Kastelein RA,Kumar S,Lin H,Mulero JJ,Pan J,Pan Y,Smith DE,Young PR(2001)."A new nomenclature for IL-1-family genes".Trends Immunol.22(10):536-7.
[35]Dunn E,Sims JE,Nicklin MJ,O'Neill LA(2001)."Annotating genes with potential roles in the immune system:six new members of the IL-1 family".Trends Immunol.22(10):533-6.
[36]Arend WP,Palmer G,Gabay C.IL-1,IL-18,and IL-33 families of cytokines.Immunol Rev 2008 Jun;223:20-38.
[37]Radons J,Gabler S,Wesche H,Korherr C,Hofmeister R,Falk W.Identification of essential regions in the cytoplasmic tail of interleukin-1 receptor accessory protein critical for interleukin-1 signaling.J Biol Chem 2002,277:16456-16463.
[38]Wesche H,Korherr C,Kracht M,Falk W,Resch K,Martin MU(March 1997)."The interleukin-1 receptor accessory protein(IL-1RAcP) is essential for IL-1-induced activation of interleukin-1 receptor-associated kinase(IRAK) and stress-activated protein kinases(SAP kinases)".J.Biol.Chem.272(12):7727-31.
[39]Jun Zou,Charles Cunningham,and Christopher J.Secombes.The rainbow trout Oncorhynchus mykiss interleukin-1b gene has a different organization to mammals and undergoes incomplete splicing.Eur.J.Biochem.259,901±908(1999)
[40]Kumar S,Hedges SB.A molecular timescale for vertebrate evolution.Nature 1998;392(6679):917-20.
[41]郭尧君:蛋白质电泳实验技术,科学出版社;1999.
[42]Watters TM,Kenny EF,O'Neill LA.Structure,function and regulation of the Toll/IL-1 receptor adaptor proteins.Immunol Cell Biol.2007 Aug-Sep;85(6):411-9.Epub 2007 Jul 31.
[43]Hurley IA,Mueller RL,Dunn KA,Sclunidt EJ,Friedman M,Ho RK et al.A new time-scale for ray-finned fish evolution.Proc Biol Sci 2007;274(1609):489-98.
[2]Barksby HE,Lea SR,Preshaw PM,Taylor JJ:The expanding family of interleukin-1 cytokines and their role in destructive inflammatory disorders.Clin Exp Immunol 2007,149(2):217-225.
[3]Nicola,N.A.(1994).Guidebook to cytokines and their receptors.Oxford."Sambrook and Tooze.
[4]Roux-Lombard.P.The interleukin-1 family.Eur Cytokine Netw 1998,9(4):565-576.
[5]Morgan MM,Clayton CC,Heinricher MM(2004)."Dissociation of hyperalgesia from fever following intracerebroventricular administration of interleukin-lbeta in the rat".Brain Res.1022(1-2):96-100.
[6]Vigers,G.P.A.,Anderson,L.J.,Caes,P.& Brandhuber,B.J.(1997).Crystal structure of the type-1 interleukin-1 receptor complexed with interleukin-1 a.Nature 386,190-194
[7]Ellsaesser,C.F.& Clem,L.W.(1994).Functionally distinct high and low molecular weight species of channel catfish and mouse IL-1.Cytokine 6,10-20
[8]Verburg van Kemenade,B.M.L.,Weyts,F.A.A.,Debets,R.& Flik,G.(1995).Carp macrophages and neutrophilic granulocytes secrete an interleukin-1-like factor.Developmental and Comparative Immunology 19,59-70.
[9]Beuscher H,Gunther C,Rollinghoff M.IL-1 beta is secreted by activated murine macrophages as biologically inactive precursor.J Immunol 1990;144:2179-83.
[10]Wieczorek M.Various aspects of IL-1 biological activity.Ⅱ.IL-1 beta in diseases and the Central Nervous System.Wiad Parazytol.2009;55(2):115-24.
[11]Iizumi T.,Sato S.,Iiyama T.,Hata R.,Amemiya H.,Tomomasa H.,Yazaki T.,Umeda T.Recombinant human interleukin-1 beta analogue as a regulator of hematopoiesis in patients receiving chemotherapy for urogenital cancers.Cancer 1991;68:1520-1523
[12]Anju S.Moolwaney and Orisa J.Igwe.Regulation of the cyclooxygenase-2system by interleukin-1β through mitogen-activated protein kinase signaling pathways:A comparative study of human neuroglioma and neuroblastoma cells.Brain Res Mol Brain Res,2005,137(1-2):202-212.
[13]Javier Duque,ManuelD.Diaz-Munoz,Manuel Fresno and Miguel A.Iniguez.Up-regulation of cyclooxygenase-2 by interleukin-1β in colon carcinoma cells.Cellular Signal,2006,18(8),1262-1269.
[14]Partha Mukhopadhyay,M.Aktar Ali,Animesh Nandi,Peter Carreon,Hak Choy and Debabrata Saha.The Cyclin-Dependent Kinase 2 Inhibitor Down-regulates Interleukin-1β-Mediated Induction of Cyclooxygenase-2 Expression in Human Lung.Carcinoma Cells Cancer Res 2006;66(3):1758-66.
[15]Yun-Jin Jung,Jennifer S.Isaacs,Sunmin Lee,Jane Trepel,and Len Neckers.IL-1β mediated up-regulation of HIF-1α,via an NFkB/COX-2 pathway identifies HIF-1 as a critical link between inflammation and oncogenesis.The FASEB Journal Express Article 2003,17(14):2115-211.
[16]Gabay,C.,Porter,B.,Fantuzzi,G.& Arend,W.P.(1997).Mouse IL-1receptor antagonist isoforms:complementary DNA cloning and protein expression of intracellular isoform and tissue distribution of secreted and intracellular IL-1 receptor antagonist in vivo.Journal of Immunology 159,5905-5913.
[17]Hughes,A.L.(1994).Evolution of the interleukin-1 gene family in mammals.Journal of Molecular Evolution 39,6-12.
[18]Reddy,S A;Huang J H,Liao W S(Nov.1997)."Phosphatidylinositol 3-kinase in interleukin 1 signaling.Physical interaction with the interleukin 1 receptor and requirement in NFkappaB and AP-1 activation".J.Biol.Chem.(UNITED STATES) 272(46):29167-73.
[19]Burns,K;Clatworthy J,Martin L,Martinon F,Plumpton C,Maschera B, Lewis A,Ray K,Tschopp J,Volpe F(Jun.2000)."Tollip,a new component of the IL-1RI pathway,links IRAK to the IL-1 receptor".Nat.Cell Biol.(ENGLAND) 2(6):346-51.
[20]Muzio,M;Ni J,Feng P,Dixit V M(Nov.1997)."IRAK(Pelle) family member IRAK-2 and MyD88 as proximal mediators of IL-1 signaling".Science (UNITED STATES) 278(5343):1612-5.
[21]Huang,J;Gao X,Li S,Cao Z(Nov.1997)."Recruitment of IRAK to the interleukin 1 receptor complex requires interleukin 1 receptor accessory protein".Proc.Natl.Acad.Sci.U.S.A.(UNITED STATES) 94(24):12829-32.
[22]Copeland NG,Silan CM,Kingsley DM,et al.(1991)."Chromosomal location of murine and human IL-1 receptor genes.".Genomics 9(1):44-50.
[23]Dale M,Nicklin MJ(1999)."Interleukin-1 receptor cluster:gene organization of IL1R2,IL1R1,IL1RL2(IL-1Rrp2),IL1RL1(T1/ST2),and IL18R1(IL-1Rrp) on human chromosome 2q.".Genomics 57(1):177-9.
[24]Sims JE,Acres RB,Grubin CE,et al.(1989)."Cloning the interleukin 1receptor from human T cells.".Proc.Natl.Acad.Sci.U.S.A.86(22):8946-50.
[25]Muzio M,Ni J,Feng P,Dixit VM(1997)."IRAK(Pelle) family member IRAK-2 and MyD88 as proximal mediators of IL-1 signaling.".Science 278(5343):1612-5.
[26]Burns K,Clatworthy J,Martin L,et al.(2000)."Tollip,a new component of the IL-1RI pathway,links IRAK to the IL-1 receptor.".Nat.Cell Biol.2(6):346-51.
[27]Reddy SA,Huang JH,Liao WS(1997).Phosphatidylinositol 3-kinase in interleukin 1 signaling.Physical interaction with the interleukin 1 receptor and requirement in NFkappaB and AP-1 activation
[28]Sizemore N,Leung S,Stark GR(1999)."Activation of phosphatidylinositol 3-kinase in response to interleukin-1 leads to phosphorylation and activation of the NF-kappaB p65/RelA subunit.".Mol.Cell.Biol.19(7):4798-805.
[29]N.Morton.Interleukin-1β activates a short STAT-3 isoform in clonal insulin-secreting cells.FEBS Letters,Volume 442,Issue 1,Pages 57-60
[30]Dinarello CA.Interleukin-1,interleukin-1 receptors and interleukin-1receptor antagonist.Int Rev Immunol 1998,16(5-6):457-499.
[31]C.A.Dinarello.The interleukin-1 family:10 years of discovery.FASEB Journal.Volume 8,Issue 15,1994,pages 1314-25.
[32]Arend WP,Malyak M,Guthridge CJ,Gabay C(1998)."Interleukin-1receptor antagonist:role in biology.".Annu.Rev.Immunol.16:27-55.
[33]Huising MO,Stet RJ,Savelkoul HF,Verburg-van Kemenade BM(2004)."The molecular evolution of the interleukin-1 family of cytokines;IL-18 in teleost fish".Dev.Comp.Immunol.28(5):395-413.
[34]Sims JE,Nicklin MJ,Bazan JF,Barton JL,Busfield SJ,Ford JE,Kastelein RA,Kumar S,Lin H,Mulero JJ,Pan J,Pan Y,Smith DE,Young PR(2001)."A new nomenclature for IL-1-family genes".Trends Immunol.22(10):536-7.
[35]Dunn E,Sims JE,Nicklin MJ,O'Neill LA(2001)."Annotating genes with potential roles in the immune system:six new members of the IL-1 family".Trends Immunol.22(10):533-6.
[36]Arend WP,Palmer G,Gabay C.IL-1,IL-18,and IL-33 families of cytokines.Immunol Rev 2008 Jun;223:20-38.
[37]Radons J,Gabler S,Wesche H,Korherr C,Hofmeister R,Falk W.Identification of essential regions in the cytoplasmic tail of interleukin-1 receptor accessory protein critical for interleukin-1 signaling.J Biol Chem 2002,277:16456-16463.
[38]Wesche H,Korherr C,Kracht M,Falk W,Resch K,Martin MU(March 1997)."The interleukin-1 receptor accessory protein(IL-1RAcP) is essential for IL-1-induced activation of interleukin-1 receptor-associated kinase(IRAK) and stress-activated protein kinases(SAP kinases)".J.Biol.Chem.272(12):7727-31.
[39]Jun Zou,Charles Cunningham,and Christopher J.Secombes.The rainbow trout Oncorhynchus mykiss interleukin-1b gene has a different organization to mammals and undergoes incomplete splicing.Eur.J.Biochem.259,901±908(1999)
[40]Kumar S,Hedges SB.A molecular timescale for vertebrate evolution.Nature 1998;392(6679):917-20.
[41]郭尧君:蛋白质电泳实验技术,科学出版社;1999.
[42]Watters TM,Kenny EF,O'Neill LA.Structure,function and regulation of the Toll/IL-1 receptor adaptor proteins.Immunol Cell Biol.2007 Aug-Sep;85(6):411-9.Epub 2007 Jul 31.
[43]Hurley IA,Mueller RL,Dunn KA,Sclunidt EJ,Friedman M,Ho RK et al.A new time-scale for ray-finned fish evolution.Proc Biol Sci 2007;274(1609):489-98.