牦牛RPS5基因的克隆与原核表达
摘要
论文以我国高原牦牛为研究对象,根据GenBank中收录的普通牛RPS5基因序列(登录号为XM-589792.3)设计一对特异性引物。通过RT-PCR技术从牦牛肝脏组织总RNA中扩增出RPS5基因的编码序列,将其连接至PMD18-T载体,转化JM109感受态细胞,获得阳性重组质粒;对其进行序列测定并利用分子生物学软件进行分析,同时构建RPS5蛋白物种进化树。将牦牛RPS5基因编码区连接至Pet28a表达载体,成功构建了原核表达载体Pet28a/ Rps5;通过IPTG诱导,摸索出了融合蛋白最佳表达条件。研究结果显示,经克隆获得牦牛RPS5基因1150 bp的cDNA序列,含615 bp的开放阅读框,编码204个氨基酸;对氨基酸序列进行信号肽分析。表明该蛋白质不存在信号肽,不属于分泌性蛋白质;与已知普通牛的RPS5基因编码区进行序列比对,发现存在1个碱基的变异,但未引起氨基酸的改变,不同物种RPS5蛋白有较高的保守性;RPS5蛋白物种进化树符合物种进化规律;SDS-PAGE电泳显示在27 kD处有特异性蛋白条带,表明牦牛RPS5基因成功实现了原核表达。
This Gaoyuan Yak was takan as the candidates for the reasch. A pair of primers was designed and synthesized according to the bovine RPS5 gene sequence in Genbank (Accession Number: XM-589792.3). Coding sequence of RPS5 gene was amplified by RT-PCR technology from total RNA of yak’s liver, the product was cloned to PMD18-T vector and transfer into competent JM109-cells then got the positive recombinant; tested the sequence and analysed with molecular biology software. Phylogenetic tree of RPS5 protein was constructed. The coding region of Yak’s RPS5 gene was cloned to PET28A vector to construct an expression plasmid PET28A/ RPS5. After inducation with IPTG, it groped the best expression conditions of the fusion protein PET28A/ RPS5.
The results showed that there was a 1150bp cDNA sequence of RPS5 gene cloned, including 615 bp open reading frame, encoding 204 amino acids. This protein is not secreted protein because we did not find singal peptide via the analysis of amino acid sequences Sequence alignment indicated that there were one mutations in the coding region between yak and bovine, but don’t cause amino acid mutation in RPS5 protein. RPS5 protein has high conservation in different species; phylogenetic tree conformed to the laws of species evolution. SDS-PAGE electrophoresis showed that the band with 27 kD of molecular weight was consistent as expected. The result indicated that the successful expression of RPS5 gene。
This Gaoyuan Yak was takan as the candidates for the reasch. A pair of primers was designed and synthesized according to the bovine RPS5 gene sequence in Genbank (Accession Number: XM-589792.3). Coding sequence of RPS5 gene was amplified by RT-PCR technology from total RNA of yak’s liver, the product was cloned to PMD18-T vector and transfer into competent JM109-cells then got the positive recombinant; tested the sequence and analysed with molecular biology software. Phylogenetic tree of RPS5 protein was constructed. The coding region of Yak’s RPS5 gene was cloned to PET28A vector to construct an expression plasmid PET28A/ RPS5. After inducation with IPTG, it groped the best expression conditions of the fusion protein PET28A/ RPS5.
The results showed that there was a 1150bp cDNA sequence of RPS5 gene cloned, including 615 bp open reading frame, encoding 204 amino acids. This protein is not secreted protein because we did not find singal peptide via the analysis of amino acid sequences Sequence alignment indicated that there were one mutations in the coding region between yak and bovine, but don’t cause amino acid mutation in RPS5 protein. RPS5 protein has high conservation in different species; phylogenetic tree conformed to the laws of species evolution. SDS-PAGE electrophoresis showed that the band with 27 kD of molecular weight was consistent as expected. The result indicated that the successful expression of RPS5 gene。
引文
[1 ] Lee H S , Mun J H , Kim S G. Characterization of cDNAEncoding Cytoplasmic Ribosomal Protein L15 and L27ain Petunia ( Petunia Hybrida) : Primary Structures andCoordinate Expression[J ] . Gene , 1999 , 226 :1552163.
[2 ] van Beekvelt C A , de Graaff2Vincent M , Faber A W, etal . All three Functional Domains of the Large RibosomalSubunit Protein L25 are Required for Both Early and LatePre2rRNA Processing Steps in Saccharomyces Cerevisiae[J ] . N ucleic Acids Res , 2001 , 29 (24) : 500125008.
[3 ] Wang Q , Yang C , Zhou J , et al . Cloning and Charac2terization of Full2Length Human Ribosomal Protein L15cDNA Which was Overexpressed in Esophageal Cancer[J ] . Gene , 2001 , 263 (122) : 2052209.
[4 ] Wool I G. Extraribosomal Functions of Ribosomal Proteins[J ]. Trends Biochem Sci , 1996 , 21 : 1642165.
[5 ] Chen F W, Ioannou YA. Ribosomal Protein in Cell Pro2liferation and Apoptosis [J ] . Int Rev Immunol , 1999 ,18 : 4292448.
[6 ] Naora H, Takai I , Adach M. Altered Cellular Responses byVarying Expression of a Ribosomal Protein Gene : SequentialCoordination of Enhancement and Suppression of Ribosomal Protein S3a Gene Expression Induce Apoptosis [J ]. J CellBiol , 1998 , 141 : 7352741.
[7 ] Naora H. Involvement of Ribosomal Protein in Regulat2ing Cell Growth and Apoptosis : Translational Modulationor Recruitment for Extraribosomal Activity [ J ] . Im2munol Cell Biol , 1999 , 77 : 1972205.
[8 ] Tom M , Waterman M R , Bocking D , et al . Cloningand Characterization of the L15 Ribosomal Protein GeneHomologue from the Crayfish Orconectes Limosus [J ] .Biochem Biophys Res Commun , 1999 , 256 : 3132316.
[9 ] Song Ping , Hu Yin2chang , Xiang Zhu , et al . Cloningand Sequencing of Full2Length cDNA in Silurus Merid2ionalis Growth Hormone[J ] . Acta Hydrobiological Sini2ca , 2002 , 26 (3) : 2722280 (Ch) .
[10 ] Chomczynski P , Sacchi N. Single2Step Method of RNAIsolation by Acid Guanidinium Thiocyanate Phenol Chlo2roform Extraction [J ] . A nal Biochem , 1987 , 162 : 1562159.
[11 ] Higgins D G, Sharp P M. Fast and Sensitive Multiple Se2quence Alignments on a Microcomputer [J ] . CABIOS ,1989 , 5 (2) : 1512153.
[12 ] Kumar S , Tamura K, Jakobsen I B , et al . MEGA2 :Molecular Evolutionary Genetics Analysis software [ J ] .Bioinf ormatics , 2001 ,17 :124421245.
[13 ] Cheng Qing2tai , Zheng Bao2shan. S ystematic Search ofChinese Fishes[M] . Beijing : Science Press , 1987 , 2112352.
[14 ] Yue Pei2qi. Fauna Sinica Ostichthys : Cyprinif ormes[M] . Beijing : Science Press , 2000 , 1522488.
[15 ] Strickberge r M W. Evolution ( Third Edition ) [ M ].Boston :Jones and Bartlett Publishers , 2000. 3912461.
[16]Fisher, E. M. C, Beer- Rornero, P, Brown,L. G, Ridley, A, McNeil, J. A,Lawrence, J.B,Williard, H.F, Bieber,F.R.and Page, D.C.(1990)
[17]陈志强,徐文严.细胞凋亡与皮肤和皮肤病.国外医学皮肤性病学分册.1995, 21( 4) : 200～204
[18]曹卫红,杨志祥等.急性放射性皮肤溃疡中凋亡细胞及其相关基因变化的意义.中华放射医学与防护杂志.2003, 23( 3) : 184～186
[19]赖克方.细胞凋亡在炎症消退中的作用.国外医学呼吸系统分册,1999,19 (3): 119～121
[20]Shogo Tsuyuki ,Clande Bertrand ,FrancoisErand ,Activation of the Fas receptor on lungeosinophils leads to apoptosis and the resolutionof eosinophilic inflammation of the airways,J.Clin. Invest,1995,96:2924～2931.
[21]严威,付少颖.细胞凋亡的研究,黑龙江医学, 2002, 26( 5) : 418～450
[22] Commo S, Gaillard O, Bernard BA. Humanhair greying is linked to a specific depletionof hair follicle melanocytes affectingboth the bulb and the outer root sheath. Br JDermatol.,2004,150(3):435～443
[23]HardyM.H., The secret life of the hair follicle.Trends Genet,1992(8):55～61
[24]Sharov A, Tobi D.J., Sharov T.Y. et al.Changes in different melanocyte populationsduring hair follicle involution( catagen) .J.Znvest Dermatol,2005,125(6):1259～1267
[25]Vizirianakis IS, Pappas IS, Gougoumas D,et al. Ex pression of ribosomal protein S5cloned gene during differentiation and apoptosisin murine erythroleukemia (MEL) cells.Oncol Res. 1999;11(9):409～419
[26] Wilson KS , Kimura M, Dijk J Study on a sequence similarity be2tween ribosomal protein S5 and DNA binding proteinⅡ[J ] . FEBSLett , 1985 ,182 (2) :2492252. [27 ] Dmitry E Agafonov , Vyacheslav A Kolb , Alexander S Spirin , etal . Proteins on ribosome surface : Measurements of protein exposureby hot tritium bombardment technique [J ] . Acad Sci USA , 1997(94) :12 892212 897.
[28]杨昌山.新发现和正在研究中的细胞凋亡蛋白抑制剂[J ] .暨南大学学报:医学版,2002 ,23 (4) :20223. [29 ] Vincent Cryns ,Junying Yuan. proteases to die for[J ] . Genes &De2 velopment ,1998 ,12 :1 55121 570.
[30] Green D R ,Reed I C.Mitochondria and apoptosis[J ] . Cell ,1998 ,94 :69251.
[31] A shkennzi A ,Dixit V A. Death receptorssignaling and modulation[J ] . Science ,1998 ,281 :13 051. [32 ] Botchkareva N V ,Botchkarev V A1New roles for glial cell line2de2rived neurotrophic factor and neurotrosis : involvement in hair cyclecontrol [J ]1AmJ Pathol , 2000 ,156 (3) :1 04121 0531 [33 ] Weedon D , Strutton G1Apoptosis as the mechanism of the involu2tion of hair follicles in catagen transformation [ J ] 1Acta DermVenereol , 1981 ,61 (4) :33523391 [ 34 ] Paus R ,MulIer2Rover S1 Chronobiology of the hair follicle : huntingthe hair cycle clock[J ]1 J Investig Dermatol Symp Proc , 1999 ,4(3) :33823451
[35] Seiberg M, Jeffrey Marthinuss. Clusterin expression within skincorrelates with hair growth [J ] ,Developmental Dynamics ,1995 ,202 (3) :29423011 [36 ] Lindner G,V A Botchkarev1 Analysis of apoptosis during hair fol2 licle regression ( catagen) [ J ] 1 American Journal of Pathology ,1997 ,151 :1 60121 6171
[37] Soma T,Ogo M,Suzuki J , et al1 Analysis of apoptotic cell deathin human hair follicles in vivo. and in vitro[J ]1 Journal of Inves2 tigative Dermatology ,1998 ,111 (6) :94829521
[38] Ahmad W,Zlotogorski A ,Panteleyev A A , et al1 Genomic organi2zation of the human hairless gene (HR) and identification of amutation underlying congenital atrichia in an Arab palestinianfamily[J ]1 Genomics ,1999 ,56 (2) :14121481
[39] Costarells G, Sun T T, Lavker R M1 Label2retaining cells residein the bulge area of pilosebaceous unit : implications for follicularstem cells , hair cycle , and skin carcinogenesis[J ]1 Cell , 1990 ,61 :1 32921 337.
[40] Hide K, Daita N, Eiko H, et al. Differential exp ression ofribosomal p rotein in human nomal and neop lastic colorec2tum. J Histochem Cytochem, 2003, 51: 567～573.
[41] Wang H, Zhao L i2Na, L i Kai2Zong, et al. Overexp ression ofribosomal p rotein L15 is associated with cell p roliferation ingastric cancer. BMC Cancer, 2006, 6: 91.
[42] Reggen D, Fandolf P. Does ribosome translate cancer? JCancer, 2003, 3: 179～190.
[43] Josep Castellvi, Angel Garcia, Federico Rojo, et al. Phos2phorylated 4E binding p rotein 1: A hallmark of cell signalingthat correlates with survival in ovarian cancer. Cancer,2006, 107: 1801～1811.
[44] Gardner2Thope J , Ito H, Ashley SW, et al. Ribosomal p ro2tein P2: a potential molecular target for antisense therapy ofhuman malignancies. Anticancer Res, 2003, 10: 4549～4560.
[45] Koonin E V. Big time for small genomes. Genome Research , 1997 , 7 : 418～421
[46] Deckert G, Warren P V , Gaasterland T, et al . The complete genome of the hyperthermophilic bacterium Aquifex aeolicus. Nature ,1998 , 392 : 353～358
[47] Fraser C M, Casjens S , Huang W M, et al . Genomic sequence of a Lyme disease spirochete , Borrelia burgdorferi . Nature , 1997 ,390 : 580～586
[48] Fraser C M, Norris S J , Weinstock GM, et al . Complete genome sequence of Treponema pallidum , the syphilis spirochete. Science ,1998 , 281 : 375～388
[49] Blattner F R , PlunkettⅢG, Bloch C A , et al . The complete genome sequence of Escherichia coli K212. Science , 1997 , 277 : 1 453～1 462
[50] Fleischmann R D , Adams M D , White O , et al . Whole2genome random sequencing and assembly of Haemophilus influenzae Rd. Sci2ence , 1995 , 269 : 496～512
[51] Tomb J F , White O , Kerlavage A R , et al . The complete genome sequence of the gastric pathogen Helicobacter pylori . Nature , 1997 ,388 : 539～547
[52] Fraser C M, Gocayne J D , White O , et al . The minimal gene complement of Mycoplasma genitalium. Science , 1995 , 270 : 397～403
[53] Himmelreich R , Hilbert H , Plagens H , et al . Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae.Nucleic Acids Res , 1996 , 24 : 4 420～4 449
[54] Kunst F , Ogasawara N , Moszer I , et al . The complete genome sequence of the Gram2positive bacterium Bacillus subtilis . Nature ,1997 , 390 : 249～256
[55] Cole S T, Brosch R , Parkhill J , et al . Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Na2ture , 1998 , 393 : 537～544
[56] Kaneko T, Sato S , Kotani H , et al . Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strainPCC6803.Ⅱ. Sequence determination of the entire genome and assignment of potential2coding regions. J DNA Res , 1996 , 3 : 109～136
[57] Bult C J , White O , Olsen GJ , et al . Complete genome sequence of the methanogenicarchaeon , Methanococcus jannaschii . Science ,1996 , 273 : 1 058～1 073
[58] Smith D R , Doucette2StammL A , Deloughery C , et al . Complete genome sequence of Methanobacterium thermoautotrophicum delta H: functional analysis and comparative genomics. J Bacteriol , 1997 , 179 : 7 135～7 155
[59] Klenk H P , Clayton R A , Tomb J , et al . The complete genome sequence of the hyperthermophilic , sulphate2reducing archaeon Ar2chaeoglobus fulgidus . Nature , 1997 , 390 : 364～370
[60] Kawarabayasi Y, Sawada M, Horikawa H , et al . Complete sequence and gene organization of the genome of a hyperthermophilic archae2bacterium , Pyrococcus horikoshii OT3. J DNA Res , 1998 , 5 : 147～155
[61] Pearon W R. Rapid and sensitive sequence comparison with FASTP and FASTA. Methods Enzymol , 1990 , 183 : 63～9818 Sander C , Schneider R. Database of homology2derived protein structures and the structural meaning of sequence alignment . Proteins , 1991 , 9 : 56～68
[62] Woese C R. Archaebacteria. Sci Amer , 1981 , 244 : 98～125 20 Nomura M, Gourse R , Bauthman G. Regulation of the synthesis of ribosomes and ribosomal components. Ann Rev Biochem , 1984 , 53 :75～117
[63]罗辽复.生命进化的物理观[M ].上海:上海科学技术出版社, 2000. [ 64 ]贾力军,罗辽复,李炜疆. DNA碱基组成和碱基关联的进化[J ].内蒙古大学学报(自然科学版) , 1996, 27 (5) :635～641. [ 65 ] Pavlicek A , J abbari K, Paces J , et al. Sim ilar integrat ion but different stability of A lus and L IN Es in thehuman genome [J ]. Gene, 2001, 276 (1～2) : 39～45. [ 66 ] L uo L F. Info rmat ion bio logy: hypo theses on coding info rmat ion quant ity [J ]. A cta S cientiarum N atu raliumU niv ersitatis N eiM ong ol, 2006, 37 (3) : 285～294. [67 ]罗辽复.垃圾DNA与信息生物学[J ].科学, 2006, 58 (1) : 24～28.
[2 ] van Beekvelt C A , de Graaff2Vincent M , Faber A W, etal . All three Functional Domains of the Large RibosomalSubunit Protein L25 are Required for Both Early and LatePre2rRNA Processing Steps in Saccharomyces Cerevisiae[J ] . N ucleic Acids Res , 2001 , 29 (24) : 500125008.
[3 ] Wang Q , Yang C , Zhou J , et al . Cloning and Charac2terization of Full2Length Human Ribosomal Protein L15cDNA Which was Overexpressed in Esophageal Cancer[J ] . Gene , 2001 , 263 (122) : 2052209.
[4 ] Wool I G. Extraribosomal Functions of Ribosomal Proteins[J ]. Trends Biochem Sci , 1996 , 21 : 1642165.
[5 ] Chen F W, Ioannou YA. Ribosomal Protein in Cell Pro2liferation and Apoptosis [J ] . Int Rev Immunol , 1999 ,18 : 4292448.
[6 ] Naora H, Takai I , Adach M. Altered Cellular Responses byVarying Expression of a Ribosomal Protein Gene : SequentialCoordination of Enhancement and Suppression of Ribosomal Protein S3a Gene Expression Induce Apoptosis [J ]. J CellBiol , 1998 , 141 : 7352741.
[7 ] Naora H. Involvement of Ribosomal Protein in Regulat2ing Cell Growth and Apoptosis : Translational Modulationor Recruitment for Extraribosomal Activity [ J ] . Im2munol Cell Biol , 1999 , 77 : 1972205.
[8 ] Tom M , Waterman M R , Bocking D , et al . Cloningand Characterization of the L15 Ribosomal Protein GeneHomologue from the Crayfish Orconectes Limosus [J ] .Biochem Biophys Res Commun , 1999 , 256 : 3132316.
[9 ] Song Ping , Hu Yin2chang , Xiang Zhu , et al . Cloningand Sequencing of Full2Length cDNA in Silurus Merid2ionalis Growth Hormone[J ] . Acta Hydrobiological Sini2ca , 2002 , 26 (3) : 2722280 (Ch) .
[10 ] Chomczynski P , Sacchi N. Single2Step Method of RNAIsolation by Acid Guanidinium Thiocyanate Phenol Chlo2roform Extraction [J ] . A nal Biochem , 1987 , 162 : 1562159.
[11 ] Higgins D G, Sharp P M. Fast and Sensitive Multiple Se2quence Alignments on a Microcomputer [J ] . CABIOS ,1989 , 5 (2) : 1512153.
[12 ] Kumar S , Tamura K, Jakobsen I B , et al . MEGA2 :Molecular Evolutionary Genetics Analysis software [ J ] .Bioinf ormatics , 2001 ,17 :124421245.
[13 ] Cheng Qing2tai , Zheng Bao2shan. S ystematic Search ofChinese Fishes[M] . Beijing : Science Press , 1987 , 2112352.
[14 ] Yue Pei2qi. Fauna Sinica Ostichthys : Cyprinif ormes[M] . Beijing : Science Press , 2000 , 1522488.
[15 ] Strickberge r M W. Evolution ( Third Edition ) [ M ].Boston :Jones and Bartlett Publishers , 2000. 3912461.
[16]Fisher, E. M. C, Beer- Rornero, P, Brown,L. G, Ridley, A, McNeil, J. A,Lawrence, J.B,Williard, H.F, Bieber,F.R.and Page, D.C.(1990)
[17]陈志强,徐文严.细胞凋亡与皮肤和皮肤病.国外医学皮肤性病学分册.1995, 21( 4) : 200～204
[18]曹卫红,杨志祥等.急性放射性皮肤溃疡中凋亡细胞及其相关基因变化的意义.中华放射医学与防护杂志.2003, 23( 3) : 184～186
[19]赖克方.细胞凋亡在炎症消退中的作用.国外医学呼吸系统分册,1999,19 (3): 119～121
[20]Shogo Tsuyuki ,Clande Bertrand ,FrancoisErand ,Activation of the Fas receptor on lungeosinophils leads to apoptosis and the resolutionof eosinophilic inflammation of the airways,J.Clin. Invest,1995,96:2924～2931.
[21]严威,付少颖.细胞凋亡的研究,黑龙江医学, 2002, 26( 5) : 418～450
[22] Commo S, Gaillard O, Bernard BA. Humanhair greying is linked to a specific depletionof hair follicle melanocytes affectingboth the bulb and the outer root sheath. Br JDermatol.,2004,150(3):435～443
[23]HardyM.H., The secret life of the hair follicle.Trends Genet,1992(8):55～61
[24]Sharov A, Tobi D.J., Sharov T.Y. et al.Changes in different melanocyte populationsduring hair follicle involution( catagen) .J.Znvest Dermatol,2005,125(6):1259～1267
[25]Vizirianakis IS, Pappas IS, Gougoumas D,et al. Ex pression of ribosomal protein S5cloned gene during differentiation and apoptosisin murine erythroleukemia (MEL) cells.Oncol Res. 1999;11(9):409～419
[26] Wilson KS , Kimura M, Dijk J Study on a sequence similarity be2tween ribosomal protein S5 and DNA binding proteinⅡ[J ] . FEBSLett , 1985 ,182 (2) :2492252. [27 ] Dmitry E Agafonov , Vyacheslav A Kolb , Alexander S Spirin , etal . Proteins on ribosome surface : Measurements of protein exposureby hot tritium bombardment technique [J ] . Acad Sci USA , 1997(94) :12 892212 897.
[28]杨昌山.新发现和正在研究中的细胞凋亡蛋白抑制剂[J ] .暨南大学学报:医学版,2002 ,23 (4) :20223. [29 ] Vincent Cryns ,Junying Yuan. proteases to die for[J ] . Genes &De2 velopment ,1998 ,12 :1 55121 570.
[30] Green D R ,Reed I C.Mitochondria and apoptosis[J ] . Cell ,1998 ,94 :69251.
[31] A shkennzi A ,Dixit V A. Death receptorssignaling and modulation[J ] . Science ,1998 ,281 :13 051. [32 ] Botchkareva N V ,Botchkarev V A1New roles for glial cell line2de2rived neurotrophic factor and neurotrosis : involvement in hair cyclecontrol [J ]1AmJ Pathol , 2000 ,156 (3) :1 04121 0531 [33 ] Weedon D , Strutton G1Apoptosis as the mechanism of the involu2tion of hair follicles in catagen transformation [ J ] 1Acta DermVenereol , 1981 ,61 (4) :33523391 [ 34 ] Paus R ,MulIer2Rover S1 Chronobiology of the hair follicle : huntingthe hair cycle clock[J ]1 J Investig Dermatol Symp Proc , 1999 ,4(3) :33823451
[35] Seiberg M, Jeffrey Marthinuss. Clusterin expression within skincorrelates with hair growth [J ] ,Developmental Dynamics ,1995 ,202 (3) :29423011 [36 ] Lindner G,V A Botchkarev1 Analysis of apoptosis during hair fol2 licle regression ( catagen) [ J ] 1 American Journal of Pathology ,1997 ,151 :1 60121 6171
[37] Soma T,Ogo M,Suzuki J , et al1 Analysis of apoptotic cell deathin human hair follicles in vivo. and in vitro[J ]1 Journal of Inves2 tigative Dermatology ,1998 ,111 (6) :94829521
[38] Ahmad W,Zlotogorski A ,Panteleyev A A , et al1 Genomic organi2zation of the human hairless gene (HR) and identification of amutation underlying congenital atrichia in an Arab palestinianfamily[J ]1 Genomics ,1999 ,56 (2) :14121481
[39] Costarells G, Sun T T, Lavker R M1 Label2retaining cells residein the bulge area of pilosebaceous unit : implications for follicularstem cells , hair cycle , and skin carcinogenesis[J ]1 Cell , 1990 ,61 :1 32921 337.
[40] Hide K, Daita N, Eiko H, et al. Differential exp ression ofribosomal p rotein in human nomal and neop lastic colorec2tum. J Histochem Cytochem, 2003, 51: 567～573.
[41] Wang H, Zhao L i2Na, L i Kai2Zong, et al. Overexp ression ofribosomal p rotein L15 is associated with cell p roliferation ingastric cancer. BMC Cancer, 2006, 6: 91.
[42] Reggen D, Fandolf P. Does ribosome translate cancer? JCancer, 2003, 3: 179～190.
[43] Josep Castellvi, Angel Garcia, Federico Rojo, et al. Phos2phorylated 4E binding p rotein 1: A hallmark of cell signalingthat correlates with survival in ovarian cancer. Cancer,2006, 107: 1801～1811.
[44] Gardner2Thope J , Ito H, Ashley SW, et al. Ribosomal p ro2tein P2: a potential molecular target for antisense therapy ofhuman malignancies. Anticancer Res, 2003, 10: 4549～4560.
[45] Koonin E V. Big time for small genomes. Genome Research , 1997 , 7 : 418～421
[46] Deckert G, Warren P V , Gaasterland T, et al . The complete genome of the hyperthermophilic bacterium Aquifex aeolicus. Nature ,1998 , 392 : 353～358
[47] Fraser C M, Casjens S , Huang W M, et al . Genomic sequence of a Lyme disease spirochete , Borrelia burgdorferi . Nature , 1997 ,390 : 580～586
[48] Fraser C M, Norris S J , Weinstock GM, et al . Complete genome sequence of Treponema pallidum , the syphilis spirochete. Science ,1998 , 281 : 375～388
[49] Blattner F R , PlunkettⅢG, Bloch C A , et al . The complete genome sequence of Escherichia coli K212. Science , 1997 , 277 : 1 453～1 462
[50] Fleischmann R D , Adams M D , White O , et al . Whole2genome random sequencing and assembly of Haemophilus influenzae Rd. Sci2ence , 1995 , 269 : 496～512
[51] Tomb J F , White O , Kerlavage A R , et al . The complete genome sequence of the gastric pathogen Helicobacter pylori . Nature , 1997 ,388 : 539～547
[52] Fraser C M, Gocayne J D , White O , et al . The minimal gene complement of Mycoplasma genitalium. Science , 1995 , 270 : 397～403
[53] Himmelreich R , Hilbert H , Plagens H , et al . Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae.Nucleic Acids Res , 1996 , 24 : 4 420～4 449
[54] Kunst F , Ogasawara N , Moszer I , et al . The complete genome sequence of the Gram2positive bacterium Bacillus subtilis . Nature ,1997 , 390 : 249～256
[55] Cole S T, Brosch R , Parkhill J , et al . Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Na2ture , 1998 , 393 : 537～544
[56] Kaneko T, Sato S , Kotani H , et al . Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strainPCC6803.Ⅱ. Sequence determination of the entire genome and assignment of potential2coding regions. J DNA Res , 1996 , 3 : 109～136
[57] Bult C J , White O , Olsen GJ , et al . Complete genome sequence of the methanogenicarchaeon , Methanococcus jannaschii . Science ,1996 , 273 : 1 058～1 073
[58] Smith D R , Doucette2StammL A , Deloughery C , et al . Complete genome sequence of Methanobacterium thermoautotrophicum delta H: functional analysis and comparative genomics. J Bacteriol , 1997 , 179 : 7 135～7 155
[59] Klenk H P , Clayton R A , Tomb J , et al . The complete genome sequence of the hyperthermophilic , sulphate2reducing archaeon Ar2chaeoglobus fulgidus . Nature , 1997 , 390 : 364～370
[60] Kawarabayasi Y, Sawada M, Horikawa H , et al . Complete sequence and gene organization of the genome of a hyperthermophilic archae2bacterium , Pyrococcus horikoshii OT3. J DNA Res , 1998 , 5 : 147～155
[61] Pearon W R. Rapid and sensitive sequence comparison with FASTP and FASTA. Methods Enzymol , 1990 , 183 : 63～9818 Sander C , Schneider R. Database of homology2derived protein structures and the structural meaning of sequence alignment . Proteins , 1991 , 9 : 56～68
[62] Woese C R. Archaebacteria. Sci Amer , 1981 , 244 : 98～125 20 Nomura M, Gourse R , Bauthman G. Regulation of the synthesis of ribosomes and ribosomal components. Ann Rev Biochem , 1984 , 53 :75～117
[63]罗辽复.生命进化的物理观[M ].上海:上海科学技术出版社, 2000. [ 64 ]贾力军,罗辽复,李炜疆. DNA碱基组成和碱基关联的进化[J ].内蒙古大学学报(自然科学版) , 1996, 27 (5) :635～641. [ 65 ] Pavlicek A , J abbari K, Paces J , et al. Sim ilar integrat ion but different stability of A lus and L IN Es in thehuman genome [J ]. Gene, 2001, 276 (1～2) : 39～45. [ 66 ] L uo L F. Info rmat ion bio logy: hypo theses on coding info rmat ion quant ity [J ]. A cta S cientiarum N atu raliumU niv ersitatis N eiM ong ol, 2006, 37 (3) : 285～294. [67 ]罗辽复.垃圾DNA与信息生物学[J ].科学, 2006, 58 (1) : 24～28.