慢病毒载体介导的siRNA/shRNA抑制禽流感病毒转基因鸡的研究
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摘要
本实验采用RNA干涉(RNAi)的方法开展抗禽流感病毒转基因鸡的研究,首先将禽流感病毒H9N2毒株核蛋白基因(NP)插入真核表达载体pL-EGFP中EGFP的上游,获得了NP-EGFP融合基因表达载体pL-NP-EGFP作为禽流感病毒siRNA筛选质粒,继而针对AⅣ各亚型核蛋白基因(NP)的保守序列,设计合成三对shRNA,并退火接入带有PolⅢH1启动子的干涉质粒(pSUPER-basic-H1),分别命名为:pSUPER-siNP1、pSUPER-siNP2、pSUPER-siNP3。重组质粒经双酶切鉴定并测序确认。将此干涉质粒与NP基因筛选质粒pL-NP-EGFP(NP/EGFP infusion gene)共转染鸡胚成纤维细胞,较其他两个干涉质粒,pSUPER-siNP3高效抑制了NP基因的表达,为进一步确认比较此重组质粒对AⅣ的抑制,本实验进行了禽流感hemagglutination(HA)滴度和TCID_(50)测定,重复三次均获得相似结果,pSUPER-siNP3转染组H9N2的TCID_(50)分别降为:10~(-5.00)/0.1 mL;10~(-5.16)/0.1mL;10~(-5.28)/0.1mL;HA效价降为:1:4。而未转染干涉质粒禽流感病毒对照组TCID_((50)为:10~(-8.50)/0.1 mL,HA效价为:1:256。利用该高效干涉片段本实验构建了pLenti6-siNP3-EGFP慢病毒表达载体,并进一步确认比较了慢病毒表达质粒的干涉效果:pL-NPi-EGFP转染组H9N2的TCID_(50)降为:10~(-5.16)/0.1 mL,HA效价降为:1:4;而未转染干涉质粒禽流感病毒对照组H9N2的TCID_(50)为:10~(-8.33)/0.1 mL,HA效价为:1:256。最后将慢病毒表达载体与辅助质粒共转染293 FT细胞生产病毒颗粒。将病毒浓缩后胚下腔显微注射转染鸡胚,孵化率达10%(12/120),结果在检测的12只受体鸡胚胎中,有4只发生嵌合。
To construct expressing vector of shRNA in order to inhibit Influenza virus production.First pL-NP-EGFP,an enkaryotic expression vector containing NP-EGFP,was constructed by inserting NP gene of AIV strain into upstream of EGFP gene in eukaryotic expression vector pL-EGFP.Then targeting NP we constructed the plasmid containing PolⅢH1 promoter(pSUPER-H1).Three pairs of oligonucleotides comprised of 64 bases were chemically synthetized and annealed,pSUPER-H1 vectors were linearized with BglⅡand HindⅢ.The annealed oligonucleotides were inserted into downstream of H1 promoter to construct the recombinant shRNA plasmid(pSUPER-H1-siNP).The recombinant plasmids were identified by enzyme digestion.and sequence analysis.To detect the effect of inhibition,we cotransfect the recombinant plasmid and the screened plasmid pL-NP-EGFP(NP/EGFP infusion gene).The activity of the fluorescence in chicken embryo fibroblastic(CEF) cells transfected with the plasmid pSUPER-H1-siNP3 was lower than others and the nontransfectedcells,then It is confirmed by the hemagglutination(HA) titre and TCID_(50) of Influenza virus.The H9N2 TCID50 is 10~(5.00)/0.1 mL,HA is 1:4 transfected with pSUPER-siNP3 and the TCID_(50) of H9N2 control is 10~(-8.33)/0.1 mL,HA is 1:256.It showed that the recombinant plasmid could specifictly inhibit the virus production in CEF cells,then a lentiviral vector pL-NPi-EGFP was constructed and The H9N2 TCID_(50) is 10~(-5.16)/0.1 mL,HA is 1:4 inhibited by pL-NPi-EGFP and the TCID_(50) of H9N2 control is 10~(-8.33)/0.1 mL,HA is 1:256.then pLenti6-siNP-EGFP was constructed and harvested the virus by cotransfecting 293 FT cells with the vector and packaging plasmids.Lentiviruses after concentration were used to transfect chicken embryos,A total of 120 eggs were injected with lentivirus in our experiment from which 12(10%) chicken hatched.4 out of 12(33%) chickens were determined to contained vector sequences PCR using EGFP specific primers.
To construct expressing vector of shRNA in order to inhibit Influenza virus production.First pL-NP-EGFP,an enkaryotic expression vector containing NP-EGFP,was constructed by inserting NP gene of AIV strain into upstream of EGFP gene in eukaryotic expression vector pL-EGFP.Then targeting NP we constructed the plasmid containing PolⅢH1 promoter(pSUPER-H1).Three pairs of oligonucleotides comprised of 64 bases were chemically synthetized and annealed,pSUPER-H1 vectors were linearized with BglⅡand HindⅢ.The annealed oligonucleotides were inserted into downstream of H1 promoter to construct the recombinant shRNA plasmid(pSUPER-H1-siNP).The recombinant plasmids were identified by enzyme digestion.and sequence analysis.To detect the effect of inhibition,we cotransfect the recombinant plasmid and the screened plasmid pL-NP-EGFP(NP/EGFP infusion gene).The activity of the fluorescence in chicken embryo fibroblastic(CEF) cells transfected with the plasmid pSUPER-H1-siNP3 was lower than others and the nontransfectedcells,then It is confirmed by the hemagglutination(HA) titre and TCID_(50) of Influenza virus.The H9N2 TCID50 is 10~(5.00)/0.1 mL,HA is 1:4 transfected with pSUPER-siNP3 and the TCID_(50) of H9N2 control is 10~(-8.33)/0.1 mL,HA is 1:256.It showed that the recombinant plasmid could specifictly inhibit the virus production in CEF cells,then a lentiviral vector pL-NPi-EGFP was constructed and The H9N2 TCID_(50) is 10~(-5.16)/0.1 mL,HA is 1:4 inhibited by pL-NPi-EGFP and the TCID_(50) of H9N2 control is 10~(-8.33)/0.1 mL,HA is 1:256.then pLenti6-siNP-EGFP was constructed and harvested the virus by cotransfecting 293 FT cells with the vector and packaging plasmids.Lentiviruses after concentration were used to transfect chicken embryos,A total of 120 eggs were injected with lentivirus in our experiment from which 12(10%) chicken hatched.4 out of 12(33%) chickens were determined to contained vector sequences PCR using EGFP specific primers.
引文
[1]Dennis J.Alexander.How Dangerous Are Avian Influenza Viruses for Humans[J].World Poultry 2000:11-12
[2]Agami R.RNAi and related mechanisms and their potential use for therapy.Current Opinion in Chemical Biology 2002;6:829-834
[3]Alexander DJ.A review of avian influenza in different bird species.Veterinary Microbiology 2000;74:3-13
[4]Banks J,Speidel ES,Moore E,et al.Changes in the haemagglutinin and the neuraminidase genes prior to the emergence of highly pathogenic H7N1 avian influenza viruses in Italy.Archives of Virology 2001;146:963-973
[5]Bennink JR,Palmore TN.The promise of siRNAs for the treatment of influenza.Trends in Molecular Medicine 2004;10:571-574
[6]Bosselman RA,Hsu RY,Boggs T,et al.Germline transmission of exogenous genes in the chicken.Science 1989;243:533
[7]Bosselman RA,Hsu RY,Briskin MJ,et al.Transmission of exogenous genes into the chicken.J Reprod Fertil Suppl 1990;41:183-195
[8]Brown WRA,Hubbard SJ,Tickle C,Wilson SA.The chicken as a model for large-scale analysis of vertebrate gene function.Nature Reviews Genetics 2003;4:87-98
[9]Brummelkamp TR,Bernards R,Agami R.A System for Stable Expression of Short Interfering RNAs in Mammalian Cells.In;2002:550-553
[10]Burke B,Sumner S,Maitland N,Lewis CE.Macrophages in gene therapy:cellular delivery vehicles and in vivo targets.Journal of Leukocyte Biology 2002;72:417
[11]Chang IK,Yoshiki A,Kusakabe M,et al.Germ line chimera produced by transfer of cultured chick primordial germ cells.Cell Biology International 1995;19:569-576
[12]Chapman SC,Lawson A,MacArthur WC,et a.1.Ubiquitous GFP expression in transgenic chickens using a lentiviral vector.Development 2005;132:935-940
[13]Chen Z,Li Y,Krug RM.Influenza A virus NS1 protein targetspoly(A)-binding protein Ⅱ of the cellular 3'-end processing machinery.The EMBO Journal 1999;18:2273-2283
[14]Chin PS,Hoffmann E,Webby R,et al.Molecular Evolution of H6 Influenza Viruses from Poultry in Southeastern China:Prevalence of H6N1 Influenza Viruses Possessing Seven A/Hong Kong/156/97(H5N1)-Like Genes in Poultry.Journal of Virology 2002;76:507
[15]Choppin PW,Scheid A.The role of viral glycoproteins in adsorption,penetration,and pathogenicity of viruses.Rev Infect Dis 1980;2:40-61
[16]Claas ECJ,de Jong JC,van Beck R,Rimmelzwaan GF,Osterhaus A.Human influenza virus A/HongKong/156/97(H5N1) infection.Vaccine 1998;16:977-978
[17]Clark J,Whitelaw B.A future for transgenic livestock.Nature Reviews-Genetics 2003;4:825-833
[18]Colman PM,Laver WG,Varghese JN,et al.Three-dimensional structure of a complex of antibody with influenza virus neuraminidase.Nature 1987;326:358-363
[19]Divine JK,Flake N,Sheehan K,Gottlieb DI.Expression of a Novel Antigen,EEM-1,in the Mouse Embryo and Embryonic Stem Cell-Derived Embryoid Bodies.DEVELOPMENTAL DYNAMICS 2000;218:207-211
[20]Dolci S,Williams DE,Ernst MK,et al.Requirement for mast cell growth factor for primordial germ cell survival in culture.Nature 1991;352:809-811
[21]Dull T,Zufferey R,Kelly M,et al.A Third-Generation Lentivirus Vector with a Conditional Packaging System.Journal of Virology 1998;72:8463
[22]Elbashir SM,Harborth J,Lendeckel W,et al.Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.Nature 2001;411:494-498
[23]Enami K,Sato TA,Nakada S,Enami M.Influenza virus NS1 protein stimulates translation of the M1 protein.Journal of Virology 1994;68:1432
[124]Fodor E,Devenish L,Engelhardt OG,et al.Rescue of Influenza A Virus from Recombinant DNA.Journal of Virology 1999;73:9679
[125]Fortes P,Beloso A,Ortin J.Influenza virus NS1 protein inhibits pre-mRNA splicing and blocks mRNA nucleocytoplasmic transport.EMBO J 1994;13:704-712
[126]Gao P,Watanabe S,Ito T,et al.Biological Heterogeneity,Including Systemic Replication in Mice,of H5N1 Influenza A Virus Isolates from Humans in Hong Kong.Journal of Virology 1999;73:3184
[127]Ge Q.RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription.Proceedings of the National Academy of Sciences 2003;100:2718-2723
[28]Ge Q,Filip L,Bai A,et al.Inhibition of influenza virus production in virus-infected mice by RNA interference.Proceedings of the National Academy of Sciences 2004;101:8676
[29]Gordon JW,Scangos GA,Plotkin DJ,Barbosa JA,Ruddle FH.Genetic transformation of mouse embryos by microinjection of purified DNA.Proceedings of the National Academy of Sciences of the United States of America 1980;77:7380
[30]Gorman OT,Donis RO,Kawaoka Y,Webster RG.Evolution of influenza A virus PB2 genes:implications for evolution of the ribonucleoprotein complex and origin of human influenza A virus.J Virol 1990;64:4893-4902
[31]Guan Y,Shortridge KF,Krauss S,et al.H9N2 Influenza Viruses Possessing H5N1-Like Internal Genomes Continue To Circulate in Poultry in Southeastern China.Journal of Virology 2000;74:9372
[32]Guo YJ,Krauss S,Senne DA,et al.Characterization of the Pathogenicity of Members of the Newly Established H9N2 Influenza Virus Lineages in Asia.Virology 2000;267:279-288
[33]Han JY,Park TS,Hong YH,et al.Production of germline chimeras by transfer of chicken gonadal primordial germ cells maintained in vitro for an extended period.Theriogenology 2002;58:1531-1539
[34]Hay AJ,Lomniczi B,Bellamy AR,Skehel JJ.Transcription of the influenza virus genome.Virology 1977;83:337-355
[35]Hiromoto Y,Yamazaki Y,Fukushima T,et al.Evolutionary characterization of the six internal genes of H5N1 human influenza A virus.Journal of General Virology 2000;81:1293
[36]Hofmann A,Kessler B,Ewerling S,et al.Epigenetic Regulation of Lentiviral Transgene Vectors in a Large Animal Model.Molecular Therapy 2006;13:59-66
[37]Hofmann A,Kessler B,Ewerling S,et al.Efficient transgenesis in farm animals by lentiviral vectors.EMBO Reports 2003;4:1054
[38]Homme PJ,Easterday BC.Avian influenza virus infections.Ⅳ.Response of pheasants,ducks,and geese to influenza A-turkey-Wisconsin-1966 vires.Avian Dis 1970;14:285-290
[39]Horimoto T,Kawaoka Y.Pandemic Threat Posed by Avian Influenza A Viruses.Clinical Microbiology Reviews 2001;14:129
[40]Huang Y,Carmichael GG.A suboptimal 5' splice site is a cis-acting determinant of nuclear export of polyomavirus late mRNAs.Mol Cell Biol 1996;16:6046-6054
[41]Hunter CV,Tiley LS,Sang I-IM.Developments in transgenic technology:applications for medicine.Trends in Molecular Medicine 2005;11:293-298
[42]Jiang Y,Kang-zhen YU,Guo-hua D,et al,The Protection Efficacity of DNA Vaccine Encoding Hemagglutinin of H5 Subtype Avian Influenza Virus.中国 农业 科学(英文版)2004;3
[43]Kawakami K,Mizumoto K,Ishihama A,Shinozaki-Yamaguchi K,Miura K.Activation of Influenza Virus-Associated RNA Polymerase by Cap-1 Structure(m7GpppNm).Journal of Biochemistry 1985;97:655
[44]Kawamoto A,Gwon HC,Iwaguro H,et al.Therapeutic Potential of Ex Vivo Expanded Endothelial Progenitor Cells for Myocardial Ischemia.In:Am Heart Assoc;2001:634-637
[45]Kawaoka Y,Krauss S,Webster RG.Avian-to-human transmission of the PB1 gene of influenza A viruses in the 1957 and 1968 pandemics.J Virol 1989;63:4603-4608
[46]Krug RM,Soeiro R.Studies on the intranuclear localization of influenza virus-specific proteins.Virology 1975;64:378-387
[47]Lawrence CW,Braciale TJ.Activation,Differentiation,and Migration of Naive Virus-Specific CD8+ T Cells during Pulmonary Influenza Virus Infection 1.The Journal of Immunology 2004;173:1209-1218
[48]Lee CW,Song CS,Lee YJ,et al.Sequence analysis of the hemagglutinin gene of H9N2Korean avian influenza viruses and assessment of the pathogenic potential of isolate MS96.Avian Dis 2000;44:527-535
[49]Lewis PF,Emerman M.Passage through mitosis is required for oncoretroviruses but not for the human immunodeficiency virus.Journal of Virology 1994;68:510
[50]Li YC,Kong LH,Cheng BZ,Li KS.Construction of influenza virus siRNA expression vectors and their inhibitory effects on multiplication of influenza virus.Avian Dis 2005;49:562-573
[51]Lin YP,Shaw M,Gregory V,et al.Avian-to-human transmission of HgN2 subtype influenza A viruses:Relationship between H9N2 and H5N1 human isolates.Proceedings of the National Academy of Sciences 2000:160270697
[52]Lois C,Hong EJ,Pease S,Brown EJ,Baltimore D.Germline Transmission and Tissue-Specific Expression of Transgenes Delivered by Lentiviral Vectors.Science's STKE 2002;295:868
[53]Lu Y,Wambach M,Katze MG,Krug RM.Binding of the Influenza Virus NS1 Protein to Double-Stranded RNA Inhibits the Activation of the Protein Kinase That Phosphorylates the eIF-2 Translation Initiation Factor.Virology 1995;214:222-228
[54]Marion RM,Zurcher T,de la Luna S,Ortin J.Influenza virus NS1 protein interacts with viral transcription-replication complexes in vivo.Journal of General Virology 1997;78:2447-2451
[55]McGrew MJ,Sherman A,Ellard FM,et al.Efficient production of germline transgenic chickens using lentiviral vectors.EMBO Reports 2004;5:728-733
[156]Mead GC.Prospects for 'Competitive Exclusion'Treatment to Control Salmonellas and Other Foodbome Pathogens in Poultry.The Veterinary Journal 2000;159:111-123
[157]Meyer DB.THE MIGRATION OF PRIMORDIAL GERM CELLS IN THE CHICK EMBRYO.Dev Biol 1964;10:154-190
[158]Mo IP,Song CS,Kim KS,Rhee JC.An Occurrence of Non-Highly Pathogenic Avian Influenza in Korea.Avian Diseases 2003;47:379-383
[159]Mozdziak PE,Borwornpinyo S,McCoy DW,Petitte IN.Development of transgenic chickens expressing bacterial-galactosidase.DEVELOPMENTAL DYNAMICS 2003;226:439-445
[160]Naito M,Matsubara Y,Harumi T,et al.Foreign gene expression in the gonads of chimaeric chicken embryos by transfer of primordial germ cells transfected in vitro by lipofection for 24 hours.Animal Science Journal 2000;71:308-311
[61]Naito M,Tajima A,Yasuda Y,Kuwana T.Production of germline chimeric chickens,with high transmission rate of donor-derived gametes,produced by transfer of primordial germ cells.Molecular Reproduction and Development 1994;39:153-161
[62]Naldini L,Blomer U,Gallay P,et al.In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector.Science 1996;272:263
[63]Neumann G,Hughes MT,Kawaoka Y.Influenza A virus NS2 protein mediates vRNP nuclear export through NES-independent interaction with hCRM1.The EMBO Journal 2000;19:6751-6758
[64]Neumann G,Watanabe T,Ito H,et al.Generation of influenza A viruses entirely from cloned cDNAs.Proc Natl Acad Sci US A 1999;96:9345-9350
[65]Nordhaus WD.Alternative Approaches to the Political Business Cycle.Brookings Papers on Economic Activity 1989;2:1-49
[66]Odagiri T,Tobita K.Mutation in NS2,a nonstructural protein of influenza A virus,extragenically causes aberrant replication and expression of the PA gene and leads to generation of defective interfering particles.Proceedings of the National Academy of Sciences of the United States of America 1990;87:5988
[67]O'Neill RE.The influenza virus NEP(NS 2 protein) mediates the nuclear export of viral ribonucleoproteins.The EMBO Journal 1998;17:288-296
[68]Paddison PJ,Caudy AA,Bemstein E,Harmon GJ,Conklin DS.Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells.Genes &Development;16:948-958
[69]Palmiter RD,Brinster RL,Hammer RE,et al.Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes.Nature 1982;300:611-615
[70]Park TS,Han JY.Derivation and characterization of pluripotent embryonic germ cells in chicken.Molecular Reproduction and Development 2000;56:475-482
[71]Park YW,Katze MG.Translational Control by Influenza Virus.Journal of Biological Chemistry 1995;270:28433
[72]Paulson JC.Interactions of animal viruses with cell surface receptors.The Receptors 1985;2:131-219
[73]Peiris M,Yuen KY,Leung CW,et al.Human infection with influenza H9N2.Lancet 1999;354:916-917
[74]Pfeifer A.Lentiviral transgenesis.Transgenic Research 2004;13:513-522
[75]Pfeifer A,Ikawa M,Dayn Y,Verma IM.Transgenesis by lentiviral vectors:Lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos.Proceedings of the National Academy of Sciences 2002;99:2140
[76]Plotch SJ,Bouloy M,Ulmanen I,Krug RM.A unique cap(mTGpppXm)-dependent influenza virion endonuclease cleaves capped RNAs to generate the primers that initiate viral RNA transcription.Cell 1981;23:847-858
[77]Portela A.Replication of orthomyxoviruses Agustin Portela,* Thomas Zurcher,Amelia Nieto,and Juan Ortin.Advances in Virus Research 1999
[78]Qian XY,Alonso-Caplen F,Krug RM.Two functional domains of the influenza virus NS1protein are required for regulation of nuclear export of mRNA.Journal of Virology 1994;68:2433
[79]Qiu Y,Krug RM.The influenza virus NS1 protein is a poly(A)-binding protein that inhibits nuclear export of mRNAs containing poly(A).Journal of Virology 1994;68:2425
[80]Rapp JC,Harvey AJ,Speksnijder GL,Hu W,Ivarie R.Biologically Active Human Interferon α-2b Produced in the Egg White of Transgenic Hens.Transgenic Research 2003;12:569-575
[81]Rubinson DA,Dillon CP,Kwiatkowski AV,et al.A lentivirus-based system to functionally silence genes in primary mammalian cells,stem cells and transgenic mice by RNA interference.Nature Genetics 2003;33:401-406
[82]Sang H.Transgenesis sunny-side up.Nature Biotechnology 2006;24:955-956
[83]Scott BB,Lois C.Generation of tissue-specific transgenic birds with lentiviral vectors.Proceedings of the National Academy of Sciences 2005;102:16443-16447
[84]Shlomai A.Inhibition of hepatitis B virus expression and replication by RNA interference.Hepatology 2003;37:764-770
[85]Subbarao K,Chen H,Swayne D,et al.Evaluation of a Genetically Modified Reassortant H5N1 Influenza A Virus Vaccine Candidate Generated by Plasmid-Based Reverse Genetics.Virology 2003;305:192-200
[86]Subbarao K,Klimov A,Katz J,et al.Characterization of an Avian Influenza A(H5N1)Virus Isolated from a Child with a Fatal Respiratory Illness.Science 1998;279:393
[87]Talon J,Horvath CM,Polley R,et al.Activation of Interferon Regulatory Factor 3 Is Inhibited by the Influenza A Virus NS1 Protein.Journal of Virology 2000;74:7989
[88]Tompkins SM,Lo CY,Tumpey TM,Epstein SL.Protection against lethal influenza virus challenge by RNA interference in vivo.Proceedings of the National Academy of Sciences 2004;101:8682
[189]Tran TH,Nguyen TL,Nguyen TD,et al.Avian influenza A(H5NI) in 10 patients in Vietnam.N Engl J Med 2004;350:1179-1188
[190]van de Lavoir MC,Diamond JH,Leighton PA,et al.Germline transmission of genetically modified primordial germ cells.Nature 2006;441:766-769
[191]Vidaver AK,Koski RK,Van Etten JL.Bacteriophage φ6:a Lipid-Containing Virus of Pseudomonas phaseolicola.Journal of Virology 1973;11:799
[92]Vogel U,Woyciniuk P,Fischer A,Scholtissek C.Inhibitors of Cellular Methyltransferases and Protein Kinase C,Which Specifically Interfere with the Replication of an Influenza A Virus(Fowl Plague).Avian Diseases 2003;47:336-353
[93]von Itzstein M,Wu WY,Kok GB,et al.Rational design of potent sialidase-based inhibitors of influenza virus replication.Nature 363 1993;418:423
[94]Wagner R,Wolff T,Herwig A,Pleschka S,Klenk HD.Interdependence of Hemagglutinin Glycosylation and Neuraminidase as Regulators of Influenza Virus Growth:a Study by Reverse Genetics.Journal of Virology 2000;74:6316
[95]Wang W,Krug RM.U6atac snRNA,the highly divergent counterpart of U6 snRNA,is the specific target that mediates inhibition of AT-AC splicing by the influenza virus NS1 protein.RNA 1998;4:55-64
[96]Wang X,Li M,Zheng H,et al.Influenza A Virus NS1 Protein Prevents Activation of NF-kappa B and Induction of Alpha/Beta Interferon.Journal of Virology 2000;74:11566
[97]Williams N.Flu pandemic fears continue.Curr Biol 2005;15:R313-314
[98]Yasuda J,Nakada S,Kato A,Toyoda T,Ishihama A.Molecular assembly of influenza virus:association of the NS2 protein with virion matrix.Virology 1993;196:249-255
[99]Zajchowsld LD,Etches RJ.Transgenic chickens:past,present and future.Avian Poultry Biol Rev 2000;11:63-80
[100]Zebedee SL,Lamb RA.Influenza A virus M2 protein:monoclonal antibody restriction of virus growth and detection of M2 in virions.Journal of Virology 1988;62:2762
[101]Zhou NN,Shortridge KF,Claas ECJ,Krauss SL,Webster RG.Rapid Evolution of H5N1Influenza Viruses in Chickens in Hong Kong.Journal of Virology 1999;73:3366
[102]Zufferey R,Donello JE,Trono D,Hope TJ.Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element Enhances Expression of Transgenes Delivered by Retroviral Vectors.Journal of Virology 1999;73:2886
[103]陈伯伦,张泽纪,陈伟斌.禽流感研究(Ⅰ):鸡A型流感病毒分离与血清学初步鉴定.中国兽医杂志 1994:22:3-5
[104]黄楷,樊晓晖.新发高致病性禽流感病毒H5N1的致病性及可能起源.广西预防医学 2005;11
[105]卡尔尼克BW,高福,苏敬良,禽病学,北京.中国农业出版社 1999.In:P
[106]李宁,安道昌.生物医药中的转基因动物研究与开发.高技术通讯2000;10:106-110
[107]李赞东,刘春海,黄劲松,et al.鸡鸭异种间嵌合体的制备!动物学报 2002;48:543-548
[108]刘建忠,李宁.转基因动物研究新进展.生物技术通报 2000:11-14
[109]王小珂,李建凡.转基因禽蛋-新一代基因工程生物反应器.中国生物工程杂志2003:23:35-38
[110]Nasevicius A,Ekker SC.Effective targeted geneknoc'kdown'in zebrafish.Nature Genetics 2000;26:216-220
[111]Urban-Klein B,Werth S,Abuharbeid S,Czubayko F,Aigner A.RNAi-mediated gene-targeting through systemic application of polyethylenimine(PEI)-complexed siRNA in vivo.Gene Therapy 2005;12:461-466
[112]Foulkes JG,Liechtfried FE,Pieler C,Stephenson JR,Case CC.Methods of determining chemicals that modulate transcriptionally expression of genes associated with cardiovascular disease.In:US Patent 5,580,722;1996
[113]Pinkert CA.Transgenic animal technology:a laboratory handbook:Academic Press San Diego(USA);2002
[114].Hsiao K,Chapman P,Nilsen S,et al.Correlative Memory Deficits,A beta Elevation,and Amyloid Plaques in Transgenic Mice.Science 1996;274:99-103
[115]Towbin H,Staehelin T,Gordon J.Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets:procedure and some applications.Proceedings of the National Academy of Sciences of the United States of America 1979;76:4350
[116]Brinster RL,Chen HY,Trumbauer ME,Yagle MK,Palmiter RD.Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs.Proceedings of the National Academy of Sciences of the United States of America 1985;82:4438
[117]Song E,Lee SK,Wang J,et al.RNA interference targeting Fas protects mice from fulminant hepatitis.Nature Medicine 2003;9:347-351
[118]Zufferey R,Nagy D,Mandel RJ,Naldini L,Trono D.Multiply attenuated lentiviml vector achieves efficient gene delivery in vivo.Nature Biotechnology 1997;15:871-875
[119]Naldini L,Blomer U,Gage FH,Trono D,Verma IM.Efficient transfer,integration,and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector.Proc Natl Acad Sci US A 1996;93:11382-11388
[120]Hunt P,Li Y,Nichol JL,et al.Purification and biologic characterization of plasma-derived megakaryocyte growth and development factor.Blood 1995;86:540-547
[121]Chen X,He J,Chang LJ.Alteration of T cell immunity by lentiviral transduction of human monocyte-derived dendritic cells.Retrovirology 2004;1:37
[122]Tiscomia G,Mahadevan MS.Myotonic Dystrophy The Role of the CUG Triplet Repeats in Splicing of a Novel DMPK Exon and Altered Cytoplasmic DMPK mRNA Isoform Ratios.Molecular Cell 2000;5:959-967
[123]Natio M,Watanabe M,Kinutani M.Production of quail-chick chimaeras by blastodermal cell transfer.Br Poult Sci 1991;32:79-86
[124]Bergelson JM,Cunningham JA,Droguett G,et al.Isolation of a Common Receptor for Coxsackie B Viruses and Adenoviruses 2 and 5.Science 1997;275:1320-1323
[125]Han DW.Intestinal endotoxemia as a pathogenetic mechanism in liver failure.世界胃肠病学杂志(英文版)2002;8:961-965
[126]Van de Lavoir R,Follon M,Ravenscroft Ⅰ.Global process control information system and method.In:US Patent 5,408,603;1995
[127]Yamauchi T,Kamon J,Waki H,et al.The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity.Nature Medicine 2001;7:941-946
[128]Goldman RD,Shumaker DK,Erdos MR,et al.Accumulation of mutant lamin A causes progressive changes in nuclear architecture in Hutckinson-Gilford progeria syndrome.Proceedings of the National Academy of Sciences 2004;101:8963
[129]Bigger JT,Fleiss JL,Steinman RC,et al.Frequency domain measures of heart period variability and mortality after myocardial infarction.Circulation 1992;85:164-171
[130]Arai H,Hori S,Aramori I,Ohkubo H,Nakanishi S.Cloning and expression of a cDNA encoding an endothelin receptor.Nature 1990;348:730-732
[131]Hammer SM,Squires KE,Hughes MD,et al.A controlled trial of two nucleoside analogues plus indinavir in persons with human immunodeficiency virus infection and CD4 cell counts of 200 per cubic millimeter or less.N Engl J Med 1997;337:725-733
[132]Fox WJ.Contributions to a Knowledge of the Hymenoptera of Brazil,No.7(Continued).Proceedings of the Academy of Natural Sciences of Philadelphia 1899;51:433-464
[133]Bae YS,Kang SW,Seo MS,et al.Epidermal Growth Factor(EGF)-induced Generation of Hydrogen Peroxide.Journal of Biological Chemistry 2005;175:5423-5429
[134]Komori T,Yagi H,Nomura S,et al.Targeted disruption of Cbfal results in a complete lack of bone formation owing to maturational arrest of osteoblasts.Cell 1997;89:755-764
[135]Cheng SQ,Wang WL,Yan W,et al.Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721.世界胃肠病学杂志(英文版)2005;11:756-759
[136]Hutvagner G,Zamore PD.A microRNA in a Multiple-Turnover RNAi Enzyme Complex.Science 2002;297:2056
[137]Abrahamyan A,Nagy E,Golovan SP.DEVELOPMENT OF IN VITRO MODEL SYSTEM FOR RNA INTERFERENCE AGAINST AVIAN INFLUENZA VIRUS.78 thNortheastern Conference on Avian Diseases
[2]Agami R.RNAi and related mechanisms and their potential use for therapy.Current Opinion in Chemical Biology 2002;6:829-834
[3]Alexander DJ.A review of avian influenza in different bird species.Veterinary Microbiology 2000;74:3-13
[4]Banks J,Speidel ES,Moore E,et al.Changes in the haemagglutinin and the neuraminidase genes prior to the emergence of highly pathogenic H7N1 avian influenza viruses in Italy.Archives of Virology 2001;146:963-973
[5]Bennink JR,Palmore TN.The promise of siRNAs for the treatment of influenza.Trends in Molecular Medicine 2004;10:571-574
[6]Bosselman RA,Hsu RY,Boggs T,et al.Germline transmission of exogenous genes in the chicken.Science 1989;243:533
[7]Bosselman RA,Hsu RY,Briskin MJ,et al.Transmission of exogenous genes into the chicken.J Reprod Fertil Suppl 1990;41:183-195
[8]Brown WRA,Hubbard SJ,Tickle C,Wilson SA.The chicken as a model for large-scale analysis of vertebrate gene function.Nature Reviews Genetics 2003;4:87-98
[9]Brummelkamp TR,Bernards R,Agami R.A System for Stable Expression of Short Interfering RNAs in Mammalian Cells.In;2002:550-553
[10]Burke B,Sumner S,Maitland N,Lewis CE.Macrophages in gene therapy:cellular delivery vehicles and in vivo targets.Journal of Leukocyte Biology 2002;72:417
[11]Chang IK,Yoshiki A,Kusakabe M,et al.Germ line chimera produced by transfer of cultured chick primordial germ cells.Cell Biology International 1995;19:569-576
[12]Chapman SC,Lawson A,MacArthur WC,et a.1.Ubiquitous GFP expression in transgenic chickens using a lentiviral vector.Development 2005;132:935-940
[13]Chen Z,Li Y,Krug RM.Influenza A virus NS1 protein targetspoly(A)-binding protein Ⅱ of the cellular 3'-end processing machinery.The EMBO Journal 1999;18:2273-2283
[14]Chin PS,Hoffmann E,Webby R,et al.Molecular Evolution of H6 Influenza Viruses from Poultry in Southeastern China:Prevalence of H6N1 Influenza Viruses Possessing Seven A/Hong Kong/156/97(H5N1)-Like Genes in Poultry.Journal of Virology 2002;76:507
[15]Choppin PW,Scheid A.The role of viral glycoproteins in adsorption,penetration,and pathogenicity of viruses.Rev Infect Dis 1980;2:40-61
[16]Claas ECJ,de Jong JC,van Beck R,Rimmelzwaan GF,Osterhaus A.Human influenza virus A/HongKong/156/97(H5N1) infection.Vaccine 1998;16:977-978
[17]Clark J,Whitelaw B.A future for transgenic livestock.Nature Reviews-Genetics 2003;4:825-833
[18]Colman PM,Laver WG,Varghese JN,et al.Three-dimensional structure of a complex of antibody with influenza virus neuraminidase.Nature 1987;326:358-363
[19]Divine JK,Flake N,Sheehan K,Gottlieb DI.Expression of a Novel Antigen,EEM-1,in the Mouse Embryo and Embryonic Stem Cell-Derived Embryoid Bodies.DEVELOPMENTAL DYNAMICS 2000;218:207-211
[20]Dolci S,Williams DE,Ernst MK,et al.Requirement for mast cell growth factor for primordial germ cell survival in culture.Nature 1991;352:809-811
[21]Dull T,Zufferey R,Kelly M,et al.A Third-Generation Lentivirus Vector with a Conditional Packaging System.Journal of Virology 1998;72:8463
[22]Elbashir SM,Harborth J,Lendeckel W,et al.Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.Nature 2001;411:494-498
[23]Enami K,Sato TA,Nakada S,Enami M.Influenza virus NS1 protein stimulates translation of the M1 protein.Journal of Virology 1994;68:1432
[124]Fodor E,Devenish L,Engelhardt OG,et al.Rescue of Influenza A Virus from Recombinant DNA.Journal of Virology 1999;73:9679
[125]Fortes P,Beloso A,Ortin J.Influenza virus NS1 protein inhibits pre-mRNA splicing and blocks mRNA nucleocytoplasmic transport.EMBO J 1994;13:704-712
[126]Gao P,Watanabe S,Ito T,et al.Biological Heterogeneity,Including Systemic Replication in Mice,of H5N1 Influenza A Virus Isolates from Humans in Hong Kong.Journal of Virology 1999;73:3184
[127]Ge Q.RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription.Proceedings of the National Academy of Sciences 2003;100:2718-2723
[28]Ge Q,Filip L,Bai A,et al.Inhibition of influenza virus production in virus-infected mice by RNA interference.Proceedings of the National Academy of Sciences 2004;101:8676
[29]Gordon JW,Scangos GA,Plotkin DJ,Barbosa JA,Ruddle FH.Genetic transformation of mouse embryos by microinjection of purified DNA.Proceedings of the National Academy of Sciences of the United States of America 1980;77:7380
[30]Gorman OT,Donis RO,Kawaoka Y,Webster RG.Evolution of influenza A virus PB2 genes:implications for evolution of the ribonucleoprotein complex and origin of human influenza A virus.J Virol 1990;64:4893-4902
[31]Guan Y,Shortridge KF,Krauss S,et al.H9N2 Influenza Viruses Possessing H5N1-Like Internal Genomes Continue To Circulate in Poultry in Southeastern China.Journal of Virology 2000;74:9372
[32]Guo YJ,Krauss S,Senne DA,et al.Characterization of the Pathogenicity of Members of the Newly Established H9N2 Influenza Virus Lineages in Asia.Virology 2000;267:279-288
[33]Han JY,Park TS,Hong YH,et al.Production of germline chimeras by transfer of chicken gonadal primordial germ cells maintained in vitro for an extended period.Theriogenology 2002;58:1531-1539
[34]Hay AJ,Lomniczi B,Bellamy AR,Skehel JJ.Transcription of the influenza virus genome.Virology 1977;83:337-355
[35]Hiromoto Y,Yamazaki Y,Fukushima T,et al.Evolutionary characterization of the six internal genes of H5N1 human influenza A virus.Journal of General Virology 2000;81:1293
[36]Hofmann A,Kessler B,Ewerling S,et al.Epigenetic Regulation of Lentiviral Transgene Vectors in a Large Animal Model.Molecular Therapy 2006;13:59-66
[37]Hofmann A,Kessler B,Ewerling S,et al.Efficient transgenesis in farm animals by lentiviral vectors.EMBO Reports 2003;4:1054
[38]Homme PJ,Easterday BC.Avian influenza virus infections.Ⅳ.Response of pheasants,ducks,and geese to influenza A-turkey-Wisconsin-1966 vires.Avian Dis 1970;14:285-290
[39]Horimoto T,Kawaoka Y.Pandemic Threat Posed by Avian Influenza A Viruses.Clinical Microbiology Reviews 2001;14:129
[40]Huang Y,Carmichael GG.A suboptimal 5' splice site is a cis-acting determinant of nuclear export of polyomavirus late mRNAs.Mol Cell Biol 1996;16:6046-6054
[41]Hunter CV,Tiley LS,Sang I-IM.Developments in transgenic technology:applications for medicine.Trends in Molecular Medicine 2005;11:293-298
[42]Jiang Y,Kang-zhen YU,Guo-hua D,et al,The Protection Efficacity of DNA Vaccine Encoding Hemagglutinin of H5 Subtype Avian Influenza Virus.中国 农业 科学(英文版)2004;3
[43]Kawakami K,Mizumoto K,Ishihama A,Shinozaki-Yamaguchi K,Miura K.Activation of Influenza Virus-Associated RNA Polymerase by Cap-1 Structure(m7GpppNm).Journal of Biochemistry 1985;97:655
[44]Kawamoto A,Gwon HC,Iwaguro H,et al.Therapeutic Potential of Ex Vivo Expanded Endothelial Progenitor Cells for Myocardial Ischemia.In:Am Heart Assoc;2001:634-637
[45]Kawaoka Y,Krauss S,Webster RG.Avian-to-human transmission of the PB1 gene of influenza A viruses in the 1957 and 1968 pandemics.J Virol 1989;63:4603-4608
[46]Krug RM,Soeiro R.Studies on the intranuclear localization of influenza virus-specific proteins.Virology 1975;64:378-387
[47]Lawrence CW,Braciale TJ.Activation,Differentiation,and Migration of Naive Virus-Specific CD8+ T Cells during Pulmonary Influenza Virus Infection 1.The Journal of Immunology 2004;173:1209-1218
[48]Lee CW,Song CS,Lee YJ,et al.Sequence analysis of the hemagglutinin gene of H9N2Korean avian influenza viruses and assessment of the pathogenic potential of isolate MS96.Avian Dis 2000;44:527-535
[49]Lewis PF,Emerman M.Passage through mitosis is required for oncoretroviruses but not for the human immunodeficiency virus.Journal of Virology 1994;68:510
[50]Li YC,Kong LH,Cheng BZ,Li KS.Construction of influenza virus siRNA expression vectors and their inhibitory effects on multiplication of influenza virus.Avian Dis 2005;49:562-573
[51]Lin YP,Shaw M,Gregory V,et al.Avian-to-human transmission of HgN2 subtype influenza A viruses:Relationship between H9N2 and H5N1 human isolates.Proceedings of the National Academy of Sciences 2000:160270697
[52]Lois C,Hong EJ,Pease S,Brown EJ,Baltimore D.Germline Transmission and Tissue-Specific Expression of Transgenes Delivered by Lentiviral Vectors.Science's STKE 2002;295:868
[53]Lu Y,Wambach M,Katze MG,Krug RM.Binding of the Influenza Virus NS1 Protein to Double-Stranded RNA Inhibits the Activation of the Protein Kinase That Phosphorylates the eIF-2 Translation Initiation Factor.Virology 1995;214:222-228
[54]Marion RM,Zurcher T,de la Luna S,Ortin J.Influenza virus NS1 protein interacts with viral transcription-replication complexes in vivo.Journal of General Virology 1997;78:2447-2451
[55]McGrew MJ,Sherman A,Ellard FM,et al.Efficient production of germline transgenic chickens using lentiviral vectors.EMBO Reports 2004;5:728-733
[156]Mead GC.Prospects for 'Competitive Exclusion'Treatment to Control Salmonellas and Other Foodbome Pathogens in Poultry.The Veterinary Journal 2000;159:111-123
[157]Meyer DB.THE MIGRATION OF PRIMORDIAL GERM CELLS IN THE CHICK EMBRYO.Dev Biol 1964;10:154-190
[158]Mo IP,Song CS,Kim KS,Rhee JC.An Occurrence of Non-Highly Pathogenic Avian Influenza in Korea.Avian Diseases 2003;47:379-383
[159]Mozdziak PE,Borwornpinyo S,McCoy DW,Petitte IN.Development of transgenic chickens expressing bacterial-galactosidase.DEVELOPMENTAL DYNAMICS 2003;226:439-445
[160]Naito M,Matsubara Y,Harumi T,et al.Foreign gene expression in the gonads of chimaeric chicken embryos by transfer of primordial germ cells transfected in vitro by lipofection for 24 hours.Animal Science Journal 2000;71:308-311
[61]Naito M,Tajima A,Yasuda Y,Kuwana T.Production of germline chimeric chickens,with high transmission rate of donor-derived gametes,produced by transfer of primordial germ cells.Molecular Reproduction and Development 1994;39:153-161
[62]Naldini L,Blomer U,Gallay P,et al.In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector.Science 1996;272:263
[63]Neumann G,Hughes MT,Kawaoka Y.Influenza A virus NS2 protein mediates vRNP nuclear export through NES-independent interaction with hCRM1.The EMBO Journal 2000;19:6751-6758
[64]Neumann G,Watanabe T,Ito H,et al.Generation of influenza A viruses entirely from cloned cDNAs.Proc Natl Acad Sci US A 1999;96:9345-9350
[65]Nordhaus WD.Alternative Approaches to the Political Business Cycle.Brookings Papers on Economic Activity 1989;2:1-49
[66]Odagiri T,Tobita K.Mutation in NS2,a nonstructural protein of influenza A virus,extragenically causes aberrant replication and expression of the PA gene and leads to generation of defective interfering particles.Proceedings of the National Academy of Sciences of the United States of America 1990;87:5988
[67]O'Neill RE.The influenza virus NEP(NS 2 protein) mediates the nuclear export of viral ribonucleoproteins.The EMBO Journal 1998;17:288-296
[68]Paddison PJ,Caudy AA,Bemstein E,Harmon GJ,Conklin DS.Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells.Genes &Development;16:948-958
[69]Palmiter RD,Brinster RL,Hammer RE,et al.Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes.Nature 1982;300:611-615
[70]Park TS,Han JY.Derivation and characterization of pluripotent embryonic germ cells in chicken.Molecular Reproduction and Development 2000;56:475-482
[71]Park YW,Katze MG.Translational Control by Influenza Virus.Journal of Biological Chemistry 1995;270:28433
[72]Paulson JC.Interactions of animal viruses with cell surface receptors.The Receptors 1985;2:131-219
[73]Peiris M,Yuen KY,Leung CW,et al.Human infection with influenza H9N2.Lancet 1999;354:916-917
[74]Pfeifer A.Lentiviral transgenesis.Transgenic Research 2004;13:513-522
[75]Pfeifer A,Ikawa M,Dayn Y,Verma IM.Transgenesis by lentiviral vectors:Lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos.Proceedings of the National Academy of Sciences 2002;99:2140
[76]Plotch SJ,Bouloy M,Ulmanen I,Krug RM.A unique cap(mTGpppXm)-dependent influenza virion endonuclease cleaves capped RNAs to generate the primers that initiate viral RNA transcription.Cell 1981;23:847-858
[77]Portela A.Replication of orthomyxoviruses Agustin Portela,* Thomas Zurcher,Amelia Nieto,and Juan Ortin.Advances in Virus Research 1999
[78]Qian XY,Alonso-Caplen F,Krug RM.Two functional domains of the influenza virus NS1protein are required for regulation of nuclear export of mRNA.Journal of Virology 1994;68:2433
[79]Qiu Y,Krug RM.The influenza virus NS1 protein is a poly(A)-binding protein that inhibits nuclear export of mRNAs containing poly(A).Journal of Virology 1994;68:2425
[80]Rapp JC,Harvey AJ,Speksnijder GL,Hu W,Ivarie R.Biologically Active Human Interferon α-2b Produced in the Egg White of Transgenic Hens.Transgenic Research 2003;12:569-575
[81]Rubinson DA,Dillon CP,Kwiatkowski AV,et al.A lentivirus-based system to functionally silence genes in primary mammalian cells,stem cells and transgenic mice by RNA interference.Nature Genetics 2003;33:401-406
[82]Sang H.Transgenesis sunny-side up.Nature Biotechnology 2006;24:955-956
[83]Scott BB,Lois C.Generation of tissue-specific transgenic birds with lentiviral vectors.Proceedings of the National Academy of Sciences 2005;102:16443-16447
[84]Shlomai A.Inhibition of hepatitis B virus expression and replication by RNA interference.Hepatology 2003;37:764-770
[85]Subbarao K,Chen H,Swayne D,et al.Evaluation of a Genetically Modified Reassortant H5N1 Influenza A Virus Vaccine Candidate Generated by Plasmid-Based Reverse Genetics.Virology 2003;305:192-200
[86]Subbarao K,Klimov A,Katz J,et al.Characterization of an Avian Influenza A(H5N1)Virus Isolated from a Child with a Fatal Respiratory Illness.Science 1998;279:393
[87]Talon J,Horvath CM,Polley R,et al.Activation of Interferon Regulatory Factor 3 Is Inhibited by the Influenza A Virus NS1 Protein.Journal of Virology 2000;74:7989
[88]Tompkins SM,Lo CY,Tumpey TM,Epstein SL.Protection against lethal influenza virus challenge by RNA interference in vivo.Proceedings of the National Academy of Sciences 2004;101:8682
[189]Tran TH,Nguyen TL,Nguyen TD,et al.Avian influenza A(H5NI) in 10 patients in Vietnam.N Engl J Med 2004;350:1179-1188
[190]van de Lavoir MC,Diamond JH,Leighton PA,et al.Germline transmission of genetically modified primordial germ cells.Nature 2006;441:766-769
[191]Vidaver AK,Koski RK,Van Etten JL.Bacteriophage φ6:a Lipid-Containing Virus of Pseudomonas phaseolicola.Journal of Virology 1973;11:799
[92]Vogel U,Woyciniuk P,Fischer A,Scholtissek C.Inhibitors of Cellular Methyltransferases and Protein Kinase C,Which Specifically Interfere with the Replication of an Influenza A Virus(Fowl Plague).Avian Diseases 2003;47:336-353
[93]von Itzstein M,Wu WY,Kok GB,et al.Rational design of potent sialidase-based inhibitors of influenza virus replication.Nature 363 1993;418:423
[94]Wagner R,Wolff T,Herwig A,Pleschka S,Klenk HD.Interdependence of Hemagglutinin Glycosylation and Neuraminidase as Regulators of Influenza Virus Growth:a Study by Reverse Genetics.Journal of Virology 2000;74:6316
[95]Wang W,Krug RM.U6atac snRNA,the highly divergent counterpart of U6 snRNA,is the specific target that mediates inhibition of AT-AC splicing by the influenza virus NS1 protein.RNA 1998;4:55-64
[96]Wang X,Li M,Zheng H,et al.Influenza A Virus NS1 Protein Prevents Activation of NF-kappa B and Induction of Alpha/Beta Interferon.Journal of Virology 2000;74:11566
[97]Williams N.Flu pandemic fears continue.Curr Biol 2005;15:R313-314
[98]Yasuda J,Nakada S,Kato A,Toyoda T,Ishihama A.Molecular assembly of influenza virus:association of the NS2 protein with virion matrix.Virology 1993;196:249-255
[99]Zajchowsld LD,Etches RJ.Transgenic chickens:past,present and future.Avian Poultry Biol Rev 2000;11:63-80
[100]Zebedee SL,Lamb RA.Influenza A virus M2 protein:monoclonal antibody restriction of virus growth and detection of M2 in virions.Journal of Virology 1988;62:2762
[101]Zhou NN,Shortridge KF,Claas ECJ,Krauss SL,Webster RG.Rapid Evolution of H5N1Influenza Viruses in Chickens in Hong Kong.Journal of Virology 1999;73:3366
[102]Zufferey R,Donello JE,Trono D,Hope TJ.Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element Enhances Expression of Transgenes Delivered by Retroviral Vectors.Journal of Virology 1999;73:2886
[103]陈伯伦,张泽纪,陈伟斌.禽流感研究(Ⅰ):鸡A型流感病毒分离与血清学初步鉴定.中国兽医杂志 1994:22:3-5
[104]黄楷,樊晓晖.新发高致病性禽流感病毒H5N1的致病性及可能起源.广西预防医学 2005;11
[105]卡尔尼克BW,高福,苏敬良,禽病学,北京.中国农业出版社 1999.In:P
[106]李宁,安道昌.生物医药中的转基因动物研究与开发.高技术通讯2000;10:106-110
[107]李赞东,刘春海,黄劲松,et al.鸡鸭异种间嵌合体的制备!动物学报 2002;48:543-548
[108]刘建忠,李宁.转基因动物研究新进展.生物技术通报 2000:11-14
[109]王小珂,李建凡.转基因禽蛋-新一代基因工程生物反应器.中国生物工程杂志2003:23:35-38
[110]Nasevicius A,Ekker SC.Effective targeted geneknoc'kdown'in zebrafish.Nature Genetics 2000;26:216-220
[111]Urban-Klein B,Werth S,Abuharbeid S,Czubayko F,Aigner A.RNAi-mediated gene-targeting through systemic application of polyethylenimine(PEI)-complexed siRNA in vivo.Gene Therapy 2005;12:461-466
[112]Foulkes JG,Liechtfried FE,Pieler C,Stephenson JR,Case CC.Methods of determining chemicals that modulate transcriptionally expression of genes associated with cardiovascular disease.In:US Patent 5,580,722;1996
[113]Pinkert CA.Transgenic animal technology:a laboratory handbook:Academic Press San Diego(USA);2002
[114].Hsiao K,Chapman P,Nilsen S,et al.Correlative Memory Deficits,A beta Elevation,and Amyloid Plaques in Transgenic Mice.Science 1996;274:99-103
[115]Towbin H,Staehelin T,Gordon J.Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets:procedure and some applications.Proceedings of the National Academy of Sciences of the United States of America 1979;76:4350
[116]Brinster RL,Chen HY,Trumbauer ME,Yagle MK,Palmiter RD.Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs.Proceedings of the National Academy of Sciences of the United States of America 1985;82:4438
[117]Song E,Lee SK,Wang J,et al.RNA interference targeting Fas protects mice from fulminant hepatitis.Nature Medicine 2003;9:347-351
[118]Zufferey R,Nagy D,Mandel RJ,Naldini L,Trono D.Multiply attenuated lentiviml vector achieves efficient gene delivery in vivo.Nature Biotechnology 1997;15:871-875
[119]Naldini L,Blomer U,Gage FH,Trono D,Verma IM.Efficient transfer,integration,and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector.Proc Natl Acad Sci US A 1996;93:11382-11388
[120]Hunt P,Li Y,Nichol JL,et al.Purification and biologic characterization of plasma-derived megakaryocyte growth and development factor.Blood 1995;86:540-547
[121]Chen X,He J,Chang LJ.Alteration of T cell immunity by lentiviral transduction of human monocyte-derived dendritic cells.Retrovirology 2004;1:37
[122]Tiscomia G,Mahadevan MS.Myotonic Dystrophy The Role of the CUG Triplet Repeats in Splicing of a Novel DMPK Exon and Altered Cytoplasmic DMPK mRNA Isoform Ratios.Molecular Cell 2000;5:959-967
[123]Natio M,Watanabe M,Kinutani M.Production of quail-chick chimaeras by blastodermal cell transfer.Br Poult Sci 1991;32:79-86
[124]Bergelson JM,Cunningham JA,Droguett G,et al.Isolation of a Common Receptor for Coxsackie B Viruses and Adenoviruses 2 and 5.Science 1997;275:1320-1323
[125]Han DW.Intestinal endotoxemia as a pathogenetic mechanism in liver failure.世界胃肠病学杂志(英文版)2002;8:961-965
[126]Van de Lavoir R,Follon M,Ravenscroft Ⅰ.Global process control information system and method.In:US Patent 5,408,603;1995
[127]Yamauchi T,Kamon J,Waki H,et al.The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity.Nature Medicine 2001;7:941-946
[128]Goldman RD,Shumaker DK,Erdos MR,et al.Accumulation of mutant lamin A causes progressive changes in nuclear architecture in Hutckinson-Gilford progeria syndrome.Proceedings of the National Academy of Sciences 2004;101:8963
[129]Bigger JT,Fleiss JL,Steinman RC,et al.Frequency domain measures of heart period variability and mortality after myocardial infarction.Circulation 1992;85:164-171
[130]Arai H,Hori S,Aramori I,Ohkubo H,Nakanishi S.Cloning and expression of a cDNA encoding an endothelin receptor.Nature 1990;348:730-732
[131]Hammer SM,Squires KE,Hughes MD,et al.A controlled trial of two nucleoside analogues plus indinavir in persons with human immunodeficiency virus infection and CD4 cell counts of 200 per cubic millimeter or less.N Engl J Med 1997;337:725-733
[132]Fox WJ.Contributions to a Knowledge of the Hymenoptera of Brazil,No.7(Continued).Proceedings of the Academy of Natural Sciences of Philadelphia 1899;51:433-464
[133]Bae YS,Kang SW,Seo MS,et al.Epidermal Growth Factor(EGF)-induced Generation of Hydrogen Peroxide.Journal of Biological Chemistry 2005;175:5423-5429
[134]Komori T,Yagi H,Nomura S,et al.Targeted disruption of Cbfal results in a complete lack of bone formation owing to maturational arrest of osteoblasts.Cell 1997;89:755-764
[135]Cheng SQ,Wang WL,Yan W,et al.Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721.世界胃肠病学杂志(英文版)2005;11:756-759
[136]Hutvagner G,Zamore PD.A microRNA in a Multiple-Turnover RNAi Enzyme Complex.Science 2002;297:2056
[137]Abrahamyan A,Nagy E,Golovan SP.DEVELOPMENT OF IN VITRO MODEL SYSTEM FOR RNA INTERFERENCE AGAINST AVIAN INFLUENZA VIRUS.78 thNortheastern Conference on Avian Diseases