人工感染兔出血症病毒兔肝脏的蛋白质组学分析
|
摘要
兔出血症(Rabbit hemorrhagic disease, RHD)是由兔出血症病毒(Rabbit hemorrhagic diseasevirus, RHDV)引起的兔的一种急性、败血性、高度接触性传染病，本病毒属嵌杯状病毒科（Caliciviridae）。自从上世纪八十年代首次在中国报道后，现已成为兔的一种世界流行的毁灭性传染病，给国内外养兔业带来了巨大的经济损失。病毒主要侵害兔的肝脏、肺脏和脾脏等器官，尤其以肝脏病变最为严重，是RHDV增殖的主要场所。因此，通过对肝脏群体蛋白质组学的研究对提示RHDV的感染机制和致病作用具有重大意义。本研究首次应用双向凝胶电泳比较感染后肝脏表达蛋白的差异,旨在从蛋白质群体水平上分析RHDV感染后兔肝脏蛋白表达的差异，并对筛选蛋白进行了相关功能验证，为进一步从蛋白质水平研究和揭示RHDV感染机制奠定基础。
本研究利用双向凝胶电泳（2-DE）结合质谱技术对人工感染RHDV成年兔和未感染对照组兔肝脏组织的群体蛋白质表达情况进行分析。获得49个具有统计学意义的表达差异蛋白点,其中15个在感染中表达上调,34个表达下调。生物信息学分析表明这些蛋白质主要与能量代谢、物质运输有关，同时有部分属于细胞结构蛋白和其他蛋白，其中有些在病毒感染过程中起到了作用；筛选出了6种可能在病毒感染过程中起作用的差异蛋白：α1抗胰蛋白酶（Α1AT）、血清白蛋白前体（PA）、细胞色素P-450（CYP450）、纤维蛋白原（Fg）、层粘连蛋白受体（LR）和核不均一性核糖核蛋白（hnRNP）。应用荧光定量PCR技术对其中的A1AT、PA、hnRNP的转录水平进行了分析，结果表明A1AT、PA和hnRNP在RHDV感染组肝脏组织中的mRNA水平与未感染对照组的比值分别是0.217,0.299和0.829，变化趋势和其在相应蛋白质下调表达的变化是一致的。基于2-DE分析、荧光定量PCR检测和生物信息学分析，提示A1AT可能在病毒感染中挥发作用。
从兔肝脏中提取总RNA，反转录成cDNA, PCR扩增，获得长度为1171bp的A1AT基因片段，分别与pET-30a和pEGFP-N1载体相连，分别构建A1AT的原核表达载体pET-30a-A1AT和A1AT的真核表达载体pEGFP-N1-A1AT。通过IPTG诱导原核表达，得到了可溶性A1AT蛋白。通过免疫共沉淀试验、血凝试验以及GST-pulldown试验，验证A1AT与VP60的相互作用，结果表明：免疫共沉淀试验和GST-polldown试验均表明A1AT与VP60之间存在相互作用；用A1AT中和RHDV后，血凝试验显示可抑制RHDV的血凝活性。将构建的pEGFP-N1-A1AT与pcDNA3.1-VP60重组表达载体共转染HEK293T细胞，进行激光共聚焦试验，表明A1AT能够与VP60在细胞内产生共定位。
本研究通过2-DE分析结合质谱技术，获得了人工感染RHDV兔肝脏组织的差异表达蛋白质数据；生物信息学分析和转录水平分析，确定A1AT与VP60之间可能存在相互作用；通过免疫共沉淀试验、血凝试验、GST-pulldown试验和激光共聚焦技术证明了VP60可以与A1AT蛋白发生相互作用，并存在共定位，为进一步深入研究RHDV致病机理提供了参考。
Rabbit haemorrhagic disease virus (RHDV) is a calicivirus of the genus Lagovirus that causesrabbit haemorrhagic disease (RHD) in adult rabbits. First described in China in1984, the virus rapidlyspread worldwide and is nowadays considered as endemic in several countries. The liver, lung andspleen are the primary target tissues of RHDV. The major histopathological lesions found at necropsyare acute hepatitis due to liver cell loss as the result of RHDV-induced apoptosis, and splenomegaly.Therefore, to study the differences of protein expression profiles during the process of RHDV infectioncould help us to further elucidate the molecular mechanisms of pathogenicity.
In our study, we used proteomics techniques to study the global protein expression change profilesof rabbit liver following RHDV infection in vivo. Total liver proteins were extracted from mature rabbitchallenged by RHDV and the mock-infected. Differentially expressed proteins were separated bytwo-dimensional gel electrophoresis, analyzed by mass spectrometry (MALDI–TOF-TOF) and proteindatabase searching. Forty-nine significantly different protein expression spots were screened, amongwhich fifteen protein spots were identified to be up-regulated and thirty-four down-regulated in RHDVinfection. We presume that five of them were involved with the infection of RHDV, includingalpha-1-antitrypsin（A1AT），serum albumin precursor(PA), laminin receptor，cytochrome P-450，fibrinogen and heterogeneous nuclear ribonucleoprotein（hnRNP）. In addition, the transcriptional levelsof3genes (A1AT，PA and hnRNP) were verified by real time PCR, two of which was consistent withthe proteomic result. Next we also proceeded with further interaction verification and functional studiesof Α1AT which is one of the significantly different protein expression spots.
To prokaryotic and eukaryotic express the fragment of A1AT gene, the fragment were amplified byPCR with primers designed according to the sequence of A1AT gene and inserted into pET-30a vectorfor expression in E.coli and pEGFP-N1vector for confocal assay respectively. The interaction betweenA1AT protein and VP60could be verified by immunoprecipitation test and HI test. The GST-polldowntest also indicated that VP60could interact with A1AT protein. Finally, with the help of confocal lasertechnology we verified the interaction between A1AT and VP60after pEGFP-N1-Msn andpcDNA3.1-VP60co-transfected into HEK293T cells.
In summary, the proteomics analysis of rabbit Liver provides valuable insights into the interactionsof RHDV with its host and the investigations in pathogenesis of RHDV. Meanwhile, the results of theinteraction between VP60and A1AT in our studies would be facilitated further study of thepathogenesis of RHDV and Host-virus co-evolution.
本研究利用双向凝胶电泳（2-DE）结合质谱技术对人工感染RHDV成年兔和未感染对照组兔肝脏组织的群体蛋白质表达情况进行分析。获得49个具有统计学意义的表达差异蛋白点,其中15个在感染中表达上调,34个表达下调。生物信息学分析表明这些蛋白质主要与能量代谢、物质运输有关，同时有部分属于细胞结构蛋白和其他蛋白，其中有些在病毒感染过程中起到了作用；筛选出了6种可能在病毒感染过程中起作用的差异蛋白：α1抗胰蛋白酶（Α1AT）、血清白蛋白前体（PA）、细胞色素P-450（CYP450）、纤维蛋白原（Fg）、层粘连蛋白受体（LR）和核不均一性核糖核蛋白（hnRNP）。应用荧光定量PCR技术对其中的A1AT、PA、hnRNP的转录水平进行了分析，结果表明A1AT、PA和hnRNP在RHDV感染组肝脏组织中的mRNA水平与未感染对照组的比值分别是0.217,0.299和0.829，变化趋势和其在相应蛋白质下调表达的变化是一致的。基于2-DE分析、荧光定量PCR检测和生物信息学分析，提示A1AT可能在病毒感染中挥发作用。
从兔肝脏中提取总RNA，反转录成cDNA, PCR扩增，获得长度为1171bp的A1AT基因片段，分别与pET-30a和pEGFP-N1载体相连，分别构建A1AT的原核表达载体pET-30a-A1AT和A1AT的真核表达载体pEGFP-N1-A1AT。通过IPTG诱导原核表达，得到了可溶性A1AT蛋白。通过免疫共沉淀试验、血凝试验以及GST-pulldown试验，验证A1AT与VP60的相互作用，结果表明：免疫共沉淀试验和GST-polldown试验均表明A1AT与VP60之间存在相互作用；用A1AT中和RHDV后，血凝试验显示可抑制RHDV的血凝活性。将构建的pEGFP-N1-A1AT与pcDNA3.1-VP60重组表达载体共转染HEK293T细胞，进行激光共聚焦试验，表明A1AT能够与VP60在细胞内产生共定位。
本研究通过2-DE分析结合质谱技术，获得了人工感染RHDV兔肝脏组织的差异表达蛋白质数据；生物信息学分析和转录水平分析，确定A1AT与VP60之间可能存在相互作用；通过免疫共沉淀试验、血凝试验、GST-pulldown试验和激光共聚焦技术证明了VP60可以与A1AT蛋白发生相互作用，并存在共定位，为进一步深入研究RHDV致病机理提供了参考。
Rabbit haemorrhagic disease virus (RHDV) is a calicivirus of the genus Lagovirus that causesrabbit haemorrhagic disease (RHD) in adult rabbits. First described in China in1984, the virus rapidlyspread worldwide and is nowadays considered as endemic in several countries. The liver, lung andspleen are the primary target tissues of RHDV. The major histopathological lesions found at necropsyare acute hepatitis due to liver cell loss as the result of RHDV-induced apoptosis, and splenomegaly.Therefore, to study the differences of protein expression profiles during the process of RHDV infectioncould help us to further elucidate the molecular mechanisms of pathogenicity.
In our study, we used proteomics techniques to study the global protein expression change profilesof rabbit liver following RHDV infection in vivo. Total liver proteins were extracted from mature rabbitchallenged by RHDV and the mock-infected. Differentially expressed proteins were separated bytwo-dimensional gel electrophoresis, analyzed by mass spectrometry (MALDI–TOF-TOF) and proteindatabase searching. Forty-nine significantly different protein expression spots were screened, amongwhich fifteen protein spots were identified to be up-regulated and thirty-four down-regulated in RHDVinfection. We presume that five of them were involved with the infection of RHDV, includingalpha-1-antitrypsin（A1AT），serum albumin precursor(PA), laminin receptor，cytochrome P-450，fibrinogen and heterogeneous nuclear ribonucleoprotein（hnRNP）. In addition, the transcriptional levelsof3genes (A1AT，PA and hnRNP) were verified by real time PCR, two of which was consistent withthe proteomic result. Next we also proceeded with further interaction verification and functional studiesof Α1AT which is one of the significantly different protein expression spots.
To prokaryotic and eukaryotic express the fragment of A1AT gene, the fragment were amplified byPCR with primers designed according to the sequence of A1AT gene and inserted into pET-30a vectorfor expression in E.coli and pEGFP-N1vector for confocal assay respectively. The interaction betweenA1AT protein and VP60could be verified by immunoprecipitation test and HI test. The GST-polldowntest also indicated that VP60could interact with A1AT protein. Finally, with the help of confocal lasertechnology we verified the interaction between A1AT and VP60after pEGFP-N1-Msn andpcDNA3.1-VP60co-transfected into HEK293T cells.
In summary, the proteomics analysis of rabbit Liver provides valuable insights into the interactionsof RHDV with its host and the investigations in pathogenesis of RHDV. Meanwhile, the results of theinteraction between VP60and A1AT in our studies would be facilitated further study of thepathogenesis of RHDV and Host-virus co-evolution.
引文
1.陈尧,王亦男,核不均一核糖核蛋白功能与疾病关系的研究进展.生理科学进展,2008，39(2)：109～112.
2.郭春燕,詹克辉,蛋白质组学技术进展及应用.云南农业大学学报,2010,25(4):583～588
3.郭纯,免疫共沉淀技术的研究进展.中医药导报,2007，13(13)：86～88.
4.郭明章,董亚芳,罗函禄,动物血清中兔出血症病毒非特异性血凝抑制因子的研究.中国养兔杂志,1992,(1):19～22.
5.胡晓峰，刘斌，张敏,家兔肝纤维化模型的建立及肝功能指标与肝纤维化的相关性.实用肝脏病杂志，2007,10（3）150～152.
6.姜茜,贾凌云,蛋白质相互作用研究的新技术和新方法.中国生物化学与分子生物学报，2008,24(10)：974～979.
7.李德军,邓治,陈春柳,植物组织双向电泳样品制备方法研究进展.中国农学通报,2009,25(24):78～82.
8.李昊,李义,免疫共沉淀技术的研究进展.内蒙古医学杂志,2008,40(4)：452-455.
9.林治涌,兔瘟血凝及血凝抑制试验操作方法.畜牧兽医杂志，1995,14（2）：53-55
10.刘庆,李永光,纤维蛋白原在急性脑出血时的变化及作用.西部医学,2011，23(2)：252～255.
11.刘庆,李永光,炎症反应和凝血因子在急性脑血管病中的作用及应用价值.中国神经免疫学和神经病学杂志,2010，17(5)：377～380.
12.卢建红,邵卫星,龙进学, A型流感病毒毒力因子研究进展.动物医学进展.2004,25(4):63～66
13.马驰,刘淑清,宋军卫,双向电泳结合质谱初步分析蛇岛蝮蛇毒蛋白质组.动物学杂志,2009,44(4):70～77.
14.孟箐箐,黄大毛,唐发清,蛋白质与蛋白质相互作用的研究进展.国际病理科学与临川杂志,2008,28(6)：471～482.
15.莫莉,黄大毛,唐发清,分子动态模拟及其在生物大分子研究中的应用.生命的化学,2007,(3):2432～2454.
16.邱家山,赵红卫, α1-抗胰蛋白酶的分离纯化及应用研究.生命科学研究,2005,9(4):47.
17.桑志红.电喷雾电离质谱及其在蛋白质化学研究中的应用.国外医学.药学分册,2000,27(1):38.
18.盛祖恬，周明龙,兔出血热病毒提纯和多肽的比较研究.畜牧兽医学报,1994,25（4）:380-388郑东，谌东华.低温电镜术证实了兔出血症病毒亚基因组颗粒的存在.电子显微学报,2000，19：660～666.
19.宋秉生,酶联免疫吸附试验和免疫印迹技术检测兔血液滤纸片中兔出血症病毒抗体.中国兽医杂志,1996,22(11):6.
20.熊梅，穆亚芳，刘家森，与兔出血症病毒VP60蛋白相互作用蛋白的初步鉴定.中国预防兽医学报,2010，32(6)：479～481.
21.徐天祥,血清前白蛋白对肝脏疾病的诊断价值.当代医学,2009，15(16)：19～21.
22.许克新,王云川,双向电泳及质谱分析与蛋白组研究.第四军医大学学报,2002,23(22):2017～2022.
23.严维巍，朱国强,王永坤,兔出血症病毒研究进展.中国养兔,2001，1：26～29.
24.殷震，刘景华主编,动物病毒学(第二版).北京:科学出版社,1996，525～529.
25.于靖,王方,蛋白质组学研究技术及其联合应用.医学分子生物学杂志,2007,4(4):371～374.
26.于莉娜,钟雪云,陈运贤,免疫共沉淀法用于肿瘤异质性研究的可行性分析.免疫学杂志,2004,20(1):58～64.
27.袁顺宗，彭旭，马兵，利用免疫共沉淀验证HTO36与P311间的相互作用.第三军医大学学报,2006，28(24)：2400～2402.
28.袁顺宗，吴军，易绍营，以酵母双杂交系统从成人肝cDNA文库中筛选与研究P31相互作用蛋白的基因序列.第三军医大学学报,2005，27(14)：1428～1431.
29.曾群辉，朱国玉，索珍，西藏兔病毒性出血症的研究.中国预防兽医学报，2000,24（2），96～98
30.詹显全,陈主初,蛋白质组中蛋白质鉴定技术的研究近况.国外医学分子生物学分册,2002,24(3):129～133
31.赵晓芳,孟程程,李招发,层粘连蛋白与疾病相关性的研究进展.生物技术通讯,2010,21(1):116～121.
32.祝慈芳,朱威, α1-抗胰蛋白酶制剂及其防治急性肺损伤的疗效.中国生物制品学杂志,2004,17(1):39～41.
33. Ahn T., Yun CH., Molecular mechanisms regulating the mitochondrial targeting of microsomalcytochrome P450enzymes. Curr Drug Metab,2010,11(10):830-839
34. Alfonso P., Rivera J., Hernaez B., et al., Identification of cellular proteinsmodified in response toAfrican swine fever virus infection by proteomics. Proteomics2004,4:2037-2046
35. Alonso J M., R Casais, J A Boga, et al., Processing of rabbit hemorrhagic disease virus polyprotein.J Virol.1996,70:1261-1265.
36. Anderson, N. G.&Anderson, N. L. Twenty years of two-dimensional electrophoresis: past,presentand future. Electrophoresis17,443-453
37. Barcena J., Verdaguer N., Roca R., et al., The coat protein of Rabbit hemorrhagic disease viruscontains a molecular switch at the N-terminal region facing the inner surface of the capsid.Virology2004,322:118-134.
38. Betts J C., Blackstock, W P., et al., Identification of phosphorylation sites on neurofilamentproteins by nanoelectrospray mass spectrometry. J. Biol. Chem,1997,272,12922-12927.
39. Bonkovsky H L., Banner B F., Rothman A L., Iron and chronic viral hepatitis. Hepatology(Baltimore, Md.)1997,25(3):759-68.
40. Bustin S A., Absolute quantification of mRNA using real-time reverse transcription polymerasechain reaction assays. Journal of Molecular Endocrinology,2000,25:169-193.
41. Cancellotti FM., Renzi M: Epidemiology and current situation of viral haemorrhagic disease ofrabbits and the European brown hare syndrome in Italy. Rev Sci Tech1991,10:409-422.
42. Capucci L., Frigoli G., Ronshold L., et al., Antigenicity of the rabbit hemorrhagic disease virusstudied by its reactivity with monoclonal antibodies.Virus Res1995,37:221-238.
43. Capucci L., Scicluna MT., Lavazza A., Diagnosis of viral haemorrhagic disease of rabbits and theEuropean brown hare syndrome. Rev Sci Tech1991,10:347-370.
44. CAPUTI, AM ZAHLER. Determination of the RNA binding specificity of the heterogeneousnuclear ribonucleoprotein family．J Biol Chem，2001．276(47):43850-43859．
45. Carlson JA., Rogers BB., Sifers RN., et al., Multiple tissues express alpha1-antitrypsin intransgenic mice and man. J Clin Invest1988,82(1):26-36
46. Chahed K., Kabbage M., Hamrita B., et al., Detection of protein alterations in male breast cancerusing two dimensional gel electrophoresis and mass spectrometry: the involvement of severalpathways in tumorigenesis. ClinChim Acta2008,388:106-114
47. Chai J., Du C., Wu J.W., et al., Structural and biochemical basis of apoptotic activation bySmac/DIABLO. Nature,2000,406(6798),855-862.
48. Choi JM., Hutson AM., Estes MK., Atomic resolution structural characterization of recognition ofhisto-blood group antigens by Norwalk virus. Proc Natl Acad Sci USA2008,105:9175-9180.
49. Cooke BD., Rabbit haemorrhagic disease: field epidemiology and the management of wild rabbitpopulations. Rev Sci Tech2002,21:347-358.
50. D’Alession J M&Gerard G F., Second-strand cDNA synthesis with E.coli DNA polylmerase I andRNase H: the fate of information at the mRNA5’ terminus and the effect of E.coli DNA ligase.Nucleic Acid Res.1988,16:1999-2014.
51. Dalpke A.H., Thomssen R., Ritter K., Oxidative injury to endothelial cells due toEpstein-Barrvirus-induced autoantibodies against manganese superoxide dismutase. J Med Virol2003,71:408-416.
52. Delibes-Mateos M., Delibes M., Ferreras P., Villafuerte R: Key role of European rabbits in theconservation of the Western Mediterranean basin hotspot. Conserv Biol2008,22:1106-1117.
53. Dreyfuss G., Matunis M J., Pino-lRoma S., et al. HnRNP proteins and the biogenesis of mRNA.Annu Rev Bi ochem,1993,62(1):289-321
54. Eckerkorn C., et al., Mass spectrometric analysis of blotted proteins after gel electrophoresisseparation by matrix-assisted laser desorption. Electrophoresis13,1992,664-665.
55. Elliott PR., Bilton D., Lomas DA., Lung polymers in Z alpha1-antitrypsin defciency-relatedemphysema. Am J Respir Cell Mol Biol,1998,18(5):670-674
56. Esteves PJ., Abrantes J., Carneiro M., et al., Detection of positive selection in the major capsidprotein VP60of the rabbit haemorrhagic disease virus (RHDV). Virus Res2008,137:253-256.
57. Farkas T., Sestak K., Wei C., et al., Characterization of a rhesus monkey calicivirus representing anew genus of Caliciviridae. J Virol2008,82:5408-5416.
58. Farnos O., Boue O., Parra F., High-level expression and immunogenic properties of therecombinant rabbit hemorrhagic disease virus VP60capsid protein obtained in Pichia pastoris.Journal of Biotechnology.2005,117(3):215-224.
59. Farnos O., Rodriguez D., Valdes O., et al., Molecular and antigenic characterization of rabbithemorrhagic disease virus isolated in Cuba indicates a distinct antigenic subtype. Arch Virol2007,152:1215-1221.
60. Fleming LE., Oquendo S., Bean JA., et al., Pilot detection study of alpha antitrypsin def-ciency ina targeted population. Am J Med Genet,2001,103(1):69-74
61. Gibb JA., Williams JM., The rabbit in New Zealand. In The European rabbit: the history andbiology of a successful colonizer. Edited by Corbet GB, Flux JEC, Rogers PM, Arthur CP, SoriguerRC, Myers K, Parer I, Wood D, Cooke BD, Gibb JA Williams JM, Fenner F, Ris J, Thompson HV,King CM. Oxford: Oxford University Press;1994:158-200.
62. Gildea TR., Shermock KM., Singer ME., et al., Cost-effectiveness analysis of augmentationtherapy for severe al-pha1-antitrypsin defciency. Am J Respir Crit Care Med,2003,167(10):1387–1392
63. Golemis EA., Brent R., Fused moietles inhibit DNA binding by LexA derivatives. Mol Cell Biol,1992,(12):3006-3014.
64. Granzow H., Weiland F., Strebelow HG., et al., Rabbit hemorrhagic disease virus (RHDV):ultrastructure and biochemical studies of typical and core-like particles present in liverhomogenates Virus Res,1996,41:163-172.
65. Gregg DA., House C., Meyer R., et al., Viral haemorrhagic disease of rabbits in Mexico:epidemiology and viral characterization. Rev Sci Tech1991,10:435-451..
66. Harrington P.R., Lindesmith L., Yount B., et al. Binding of Norwalk virus-like particles to ABHhisto-bloodgroup antigens is blocked by antisera from infected human volunteers or experimentallyvaccinated mice. J. Virol.2002,76,12335-12343
67. Henzel W.J., Billeci T.M., et al., Identifying proteins from two-dimensional gels by molecular masssearching of peptide fragments in protein sequence databases. Proc. Natl Acad. Sci.USA90,5011-5015(1993).
68. Hofstetter H., Schambock A., VanDenBerg J., et al., Specific expression of the inserted DNAsegment from hybrid plasmids constructed by the poly(dA).poly(dT)method. Biochin Biophys Acta1976,454(3):587-591.
69. Hunt SA, Baker DW, Chin MH, et al., ACC/AHA guidelines for the evaluation and management ofchronic heart failure in the adult: executive summary. A report of the ACC/AHA task force onpractice guideline.J Am Collcardio,2001,35(2):101-2113.
70. Hutson A.M., Atmar R.L., Marcus D.M., et al., Norwalkvirus-like particle hemagglutination bybinding to h histo-blood group antigens. J. Virol.2003,77,405-415.
71. Issaq H.J., Veenstra T.D., Conrads T.P., et al., The SELDI-TOF MS approach to proteomics: proteinprofiling and biomarker identification. Biochemical and biophysical research communications2002,292:587-592.
72. Jensen P. K., et al., Probing proteomes using capillary isoelectric focusing-electrosprayionizationFourier transform ion cyclotron resonance mass spectrometry. Anal. Chem.1999,71,2076-2084.
73. Juan Barcena, Nuria Verdaguer, Ramon Roca, et al., The coat protein of Rabbit hemorrhagicdisease virus contains a molecular switch at the N-terminal region facing the inner surface of thecapsid. Virology.2004,322(1):118-134.
74. Kawai T., Takahashi K., Sato S., et al., IPS-1, an adaptor triggering RIG-I-and Mda5-mediated typeI interferon induction. Nat Immunol.2005,6(10):981-8.
75. Kiledjian M., and Dreyfuss G., Primary structure and binding activity of the hnRNP U protein:binding RNA through RGG box.1992, EMBO J.11,2655-2664.
76. Larsson C., Natural history and life expectancy in severealpha1-antitrypsin defciency. Pi Z. ActaMed Scand,1978,204(5):345-351
77. Lavazza A., Capucci L., How Many Caliciviruses are there in Rabbits? A Review on RHDV andCorrelated Viruses Lagomorph Biology. Edited by Alves PC, Ferrand N, Hackl nder K: SpringerBerlin Heidelberg;2008:263-278.
78. Lavazza A., Scicluna MT., Capucci L., Susceptibility of hares and rabbits to the European brownhare syndrome virus (EBHSV) and rabbit haemorrhagic disease virus (RHDV) under experimentalconditions. Zentralbl Veterinarmed B1996,43:401-410.
79. Le Gall-Recule G., Zwingelstein F., Laurent S., et al., Phylogenetic analysis of rabbit haemorrhagicdisease virus in France between1993and2000, and the characterisation of RHDV antigenicvariants. Arch Virol2003,148:65-81.
80. Le Gall-ReculéG., Zwingelstein F., Laurent S., et al., Phylogenetic analysis of rabbit haemorrhagicdisease virus in France between1993and2000, and the characterisation of RHDV antigenicvariants. Arch Virol2003,148:65-81.
81. L'Homme Y., Sansregret R., Plante-Fortier E., et al., Genomic characterization of swinecaliciviruses representing a new genus of Caliciviridae. Virus Genes2009,39:66-75.
82. Li, J., et al., Integration of microfabricated devices to capillary electrophoresis—electrospraymassspectrometry using a low dead volume connection: application to rapid analyses ofproteolyticdigests. Anal. Chem.1999,71,3036-3045.
83. Liu SJ, Xue HP, Pu BQ, et al., A new viral disease in rabbit. Anim Husb Vet Med1984,16:253-255.
84. Lowe GDO., Rumley A., Whincup PH., et al., Hemostatic and rheological variables and risk ofcardiovascular disease. Semin Vasc Med,2002,2:429-440.
85. Mariani TD., Halluin K., et al., The Production and Analysis of Genetically Engineered MaleSterile Plants of Maize. In: Abstr35th Ann Maize Genetics Conf. Fargo：University Press，1993,35:40-45.
86. Martinez-Torrecuadrada JL., Cortes E., Vela C., et al., Antigenic structure of the capsid protein ofrabbit haemorrhagic disease virus. J Gen Virol1998,79:1901-1909.
87. McDonald, T., Sheng, S., Stanley, B., et al., Expanding the subproteome of the inner mitochondriausing protein separation technologies: one-and two-dimensional liquid chromatography andtwo-dimensional gel electrophoresis. Mol Cell Proteomics5,2392-2411.
88. Mehdaoui S., Touze A., Laurent S., et a1., Gene transfer using recombinant rabbit hemorrhagicdisease virus capsids with genetically modified DNA encapsidation capacity by addition ofpackaging sequences from or L2protein of human papillomavirus type16. J Virol.2000,74:10332-10340.
89. Meyers G., Wirblich C., Thiel HJ., Rabbit hemorrhagic disease virus--molecular cloning andnucleotide sequencing of a calicivirus genome. Virology1991,184:664-676
90. Mitro S, Krauss H: Rabbit hemorrhagic disease: a review with special reference to its epizootiology.Eur J Epidemiol1993,9:70-78.
91. Neil J D., Nucleotide sequence of the capsid protein gene of two serotypes of San Miguel sea lionvirus: identification of conserved and non-conserved amino acid sequences among caliciviruscapsid proteins. Virus Res.1992,24(2):211-222.
92. Nuwaysir, L.&Stults, J. T., ESI mass spectrometry of phosphopeptides isolated byon-lineimmobilized metal affinity chromatography. J. Am. Soc. Mass Spectrom.1993,4,662–669.
93. O'Farrell P.H., High resolution two-dimensional electrophoresis of proteins. J Biol Chem1975,250:4007-4021
94. Ohlinger VF., Haas B., Meyers G., et al., Identification and characterization of the virus causingrabbit hemorrhagic disease. J Virol1990,64:3331-3336.
95. Okayama H.&Berg P., High efficiency cloning of full-length cDNA. Mol.Cell.Bilo.1982(2):161-170.
96. P, Devaux and D, Gerlier, Antibody cross-reactivity with CD46and lack of cell surface expressionsuggest that moesin might not mediate measles virus binding. J Virol1997,71:1679~1682.
97. Park NY., Chong CY., Kim JH., et al., An outbreak of viral haemorrhagic pneumonia (tentativename) of rabbits in Korea. J Korean Vet Med Assoc1987,23:603-610.
98. Parra F., Prieto M., Purification and characterization of a calicivirus as the causative agent of alethal hemorrhagic disease in rabbits. J Virol1990,64:4013-4015.
99. Pastorino B., Boucomont-Chapeaublanc E., Peyrefitte C.N., et al., Identification of cellularproteome modifications in response to West Nilevirus infection. Mol Cell Proteomics2009,8:1623-1637.
100. Paulo JA., Lee LS., Wu B., et al., Optimized sample preparation of endoscopic collected pancreaticfluid for SDS-PAGE analysis. Elec-trophoresis.2010;31(14):2377-87.
101. Prasad BV., Rothnagel R., Jiang X., et al., Three-dimensional structure of baculovirus-expressedNorwalk virus capsids. J Virol1994,68:5117-5125.
102. Pugnale P., Latorre P., Rossi C., et al., Real-time multiplex PCR assay to quantify hepatitis C virusRNA in peripheral blood mononuclear cells. J Virol Methods,2006,133(2):1952204.
103. Qiyan J., Xifeng J., Fei Y., et al., Short cyclic peptides derived from the C-terminal sequenceof a1-antitrypsin exhibit signifcant anti-HIV-1activity. Bioorganic&Medicinal Chemistry Letters,2012,22:2393–2395.
104. Reichel C., Sudhop T., Braun B., et al., Elevated soluble tumour necrosis factor receptor serumconcentrations and short-term mortality in liver cirrhosis without acute infections.Digestion,2000,62(1):44-51．
105. Rockey DC., Noninvasive assessment of liver fibrosis and portal hypertension with transientelastography．Gastroenterology,2008,134(1):8-14．
106. Rodak L., Smid B., Valicek L., et al., Enzyme-linked immunosorbent assay of antibodies to rabbithaemorrhagic disease virus and determination of its major structural proteins. J Gen Virol1990,71:1075-1080.
107. Roman I., Figys J., Steurs G., et al., In vitro interactions between the two mitochondrial membraneproteins VDAC and cytochrome coxidase. Biochemistry2005,44(39):13192-13201.
108. Roulston A., Marcellus R.C., Branton P.E., Viruses and apoptosis. Annu Rev Microbiol1999,53:577-628
109. Schirrmeier H., Reimann I., Kollner B., et al., Pathogenic, antigenic and molecular properties ofrabbit haemorrhagic disease virus (RHDV) isolated from vaccinated rabbits: detection andcharacterization of antigenic variants. Arch.Virol.1999,144:719-735.
110. Takahashi S., Reddy SV., Chirgwin JM., et al., Cloning and identification of annexin II as anautocrine/Paracrine factor that increases osteoclast formation and bone resorption. J Biol Chem.1994,269(46):296-702.
111. Thouvenin E., Laurent S., Madelaine MF., et al., Bivalent binding of a neutralising antibody to acalicivirus involves the torsional flexibility of the antibody hinge. J Mol Biol1997,270:238-246
112. Tong, A., Wu, L., Lin, Q., et al. Proteomic analysis of cellular protein alterations using a hepatitis Bvirus-producing cellular model. Proteomics,2008,2012-2023.
113. Torrecuadradan M J L., Cortes E., Vela C., et al., Antigenic structure of the capsid protein of rabbithemorrhagic disease virus. J Gen Virol.1998,79(Pt8):1901-1909.
114. Tsui N B., Ngek., Loym., Molecular analysis of circulating RNA in plasma. Methods Mol Biol,2006,336:123-134.
115. Ushijima H., Miyamura.T, Takeda N., Proteolytic processing of sapovirus ORF1polyprotein. JVirol.2005,79:7283-7290.
116. Valicek L., Smid B., Rodah L., et al., Electron and immunoelectron microscopy of rabbithaemorrhagic disease virus (RHDV). Arch Virol,1990,112(3-4):271-275.
117. Van Dinten L C., Johan A., Wassenaar A L M., et al., An infectious arterivirus cDNA clone:Identification of a replicase point mutation that abolishes discontinuous mRNA transcription. ProcNatl Acad Sci. USA.1997,94:991-996.
118. Vester D., Rapp E., Kluge S., et al., Virus-host cell interactions in vaccineproduction cell linesinfected with different human influenza A virus variants: a proteomic approach. Journal ofproteomics2010,73:1656-1669.
119. Wiese S., Reidegeld K.A., Meyer H.E., et al., Protein labeling by iTRAQ: a new tool forquantitative mass spectrometry in proteome research. Proteomics2007,7:340-350
120. Wirblich C., Sibilia M., Boniotti MB., et al.,3C-like protease of rabbit hemorrhagic disease virus:identification of cleavage sites in the ORF1polyprotein and analysis of cleavage specificity. JVirol.1995,69(11):7159-7168.
121. Xu WY., Viral haemorrhagic disease of rabbits in the People's Republic of China: epidemiologyand virus characterization. Rev Sci Tech1991,10:393-408.
122. Y. Doi., M. Kurita., M. Matsumoto., et al., Moesin is not a receptor for Measles Virus entry intomouse embryonic stem cells. J.Virol.1998,72(2):1586-1592.
123. Yasmin MY., Aziz B., et al., Prealbumin rather than albumin is a more sensitive indicator of acuteliver disease.Malays J Pathol,1993,15(2):147.
124. Zhao Z., Huang X., Li N., et al., Direct cloning of cell differential expression genes withfull-length by a new strategy based on the multiple rounds of long distance polymerase chainreaction and magnetic beads mediated subtraction. J Bio technol1999,73:35-41.
125. Zheng X., Hong L., Shi L., et al., Proteomics analysis of host cells infected with infectious bursaldisease virus. Mol Cell Proteomics7,2008,612-6.
2.郭春燕,詹克辉,蛋白质组学技术进展及应用.云南农业大学学报,2010,25(4):583～588
3.郭纯,免疫共沉淀技术的研究进展.中医药导报,2007，13(13)：86～88.
4.郭明章,董亚芳,罗函禄,动物血清中兔出血症病毒非特异性血凝抑制因子的研究.中国养兔杂志,1992,(1):19～22.
5.胡晓峰，刘斌，张敏,家兔肝纤维化模型的建立及肝功能指标与肝纤维化的相关性.实用肝脏病杂志，2007,10（3）150～152.
6.姜茜,贾凌云,蛋白质相互作用研究的新技术和新方法.中国生物化学与分子生物学报，2008,24(10)：974～979.
7.李德军,邓治,陈春柳,植物组织双向电泳样品制备方法研究进展.中国农学通报,2009,25(24):78～82.
8.李昊,李义,免疫共沉淀技术的研究进展.内蒙古医学杂志,2008,40(4)：452-455.
9.林治涌,兔瘟血凝及血凝抑制试验操作方法.畜牧兽医杂志，1995,14（2）：53-55
10.刘庆,李永光,纤维蛋白原在急性脑出血时的变化及作用.西部医学,2011，23(2)：252～255.
11.刘庆,李永光,炎症反应和凝血因子在急性脑血管病中的作用及应用价值.中国神经免疫学和神经病学杂志,2010，17(5)：377～380.
12.卢建红,邵卫星,龙进学, A型流感病毒毒力因子研究进展.动物医学进展.2004,25(4):63～66
13.马驰,刘淑清,宋军卫,双向电泳结合质谱初步分析蛇岛蝮蛇毒蛋白质组.动物学杂志,2009,44(4):70～77.
14.孟箐箐,黄大毛,唐发清,蛋白质与蛋白质相互作用的研究进展.国际病理科学与临川杂志,2008,28(6)：471～482.
15.莫莉,黄大毛,唐发清,分子动态模拟及其在生物大分子研究中的应用.生命的化学,2007,(3):2432～2454.
16.邱家山,赵红卫, α1-抗胰蛋白酶的分离纯化及应用研究.生命科学研究,2005,9(4):47.
17.桑志红.电喷雾电离质谱及其在蛋白质化学研究中的应用.国外医学.药学分册,2000,27(1):38.
18.盛祖恬，周明龙,兔出血热病毒提纯和多肽的比较研究.畜牧兽医学报,1994,25（4）:380-388郑东，谌东华.低温电镜术证实了兔出血症病毒亚基因组颗粒的存在.电子显微学报,2000，19：660～666.
19.宋秉生,酶联免疫吸附试验和免疫印迹技术检测兔血液滤纸片中兔出血症病毒抗体.中国兽医杂志,1996,22(11):6.
20.熊梅，穆亚芳，刘家森，与兔出血症病毒VP60蛋白相互作用蛋白的初步鉴定.中国预防兽医学报,2010，32(6)：479～481.
21.徐天祥,血清前白蛋白对肝脏疾病的诊断价值.当代医学,2009，15(16)：19～21.
22.许克新,王云川,双向电泳及质谱分析与蛋白组研究.第四军医大学学报,2002,23(22):2017～2022.
23.严维巍，朱国强,王永坤,兔出血症病毒研究进展.中国养兔,2001，1：26～29.
24.殷震，刘景华主编,动物病毒学(第二版).北京:科学出版社,1996，525～529.
25.于靖,王方,蛋白质组学研究技术及其联合应用.医学分子生物学杂志,2007,4(4):371～374.
26.于莉娜,钟雪云,陈运贤,免疫共沉淀法用于肿瘤异质性研究的可行性分析.免疫学杂志,2004,20(1):58～64.
27.袁顺宗，彭旭，马兵，利用免疫共沉淀验证HTO36与P311间的相互作用.第三军医大学学报,2006，28(24)：2400～2402.
28.袁顺宗，吴军，易绍营，以酵母双杂交系统从成人肝cDNA文库中筛选与研究P31相互作用蛋白的基因序列.第三军医大学学报,2005，27(14)：1428～1431.
29.曾群辉，朱国玉，索珍，西藏兔病毒性出血症的研究.中国预防兽医学报，2000,24（2），96～98
30.詹显全,陈主初,蛋白质组中蛋白质鉴定技术的研究近况.国外医学分子生物学分册,2002,24(3):129～133
31.赵晓芳,孟程程,李招发,层粘连蛋白与疾病相关性的研究进展.生物技术通讯,2010,21(1):116～121.
32.祝慈芳,朱威, α1-抗胰蛋白酶制剂及其防治急性肺损伤的疗效.中国生物制品学杂志,2004,17(1):39～41.
33. Ahn T., Yun CH., Molecular mechanisms regulating the mitochondrial targeting of microsomalcytochrome P450enzymes. Curr Drug Metab,2010,11(10):830-839
34. Alfonso P., Rivera J., Hernaez B., et al., Identification of cellular proteinsmodified in response toAfrican swine fever virus infection by proteomics. Proteomics2004,4:2037-2046
35. Alonso J M., R Casais, J A Boga, et al., Processing of rabbit hemorrhagic disease virus polyprotein.J Virol.1996,70:1261-1265.
36. Anderson, N. G.&Anderson, N. L. Twenty years of two-dimensional electrophoresis: past,presentand future. Electrophoresis17,443-453
37. Barcena J., Verdaguer N., Roca R., et al., The coat protein of Rabbit hemorrhagic disease viruscontains a molecular switch at the N-terminal region facing the inner surface of the capsid.Virology2004,322:118-134.
38. Betts J C., Blackstock, W P., et al., Identification of phosphorylation sites on neurofilamentproteins by nanoelectrospray mass spectrometry. J. Biol. Chem,1997,272,12922-12927.
39. Bonkovsky H L., Banner B F., Rothman A L., Iron and chronic viral hepatitis. Hepatology(Baltimore, Md.)1997,25(3):759-68.
40. Bustin S A., Absolute quantification of mRNA using real-time reverse transcription polymerasechain reaction assays. Journal of Molecular Endocrinology,2000,25:169-193.
41. Cancellotti FM., Renzi M: Epidemiology and current situation of viral haemorrhagic disease ofrabbits and the European brown hare syndrome in Italy. Rev Sci Tech1991,10:409-422.
42. Capucci L., Frigoli G., Ronshold L., et al., Antigenicity of the rabbit hemorrhagic disease virusstudied by its reactivity with monoclonal antibodies.Virus Res1995,37:221-238.
43. Capucci L., Scicluna MT., Lavazza A., Diagnosis of viral haemorrhagic disease of rabbits and theEuropean brown hare syndrome. Rev Sci Tech1991,10:347-370.
44. CAPUTI, AM ZAHLER. Determination of the RNA binding specificity of the heterogeneousnuclear ribonucleoprotein family．J Biol Chem，2001．276(47):43850-43859．
45. Carlson JA., Rogers BB., Sifers RN., et al., Multiple tissues express alpha1-antitrypsin intransgenic mice and man. J Clin Invest1988,82(1):26-36
46. Chahed K., Kabbage M., Hamrita B., et al., Detection of protein alterations in male breast cancerusing two dimensional gel electrophoresis and mass spectrometry: the involvement of severalpathways in tumorigenesis. ClinChim Acta2008,388:106-114
47. Chai J., Du C., Wu J.W., et al., Structural and biochemical basis of apoptotic activation bySmac/DIABLO. Nature,2000,406(6798),855-862.
48. Choi JM., Hutson AM., Estes MK., Atomic resolution structural characterization of recognition ofhisto-blood group antigens by Norwalk virus. Proc Natl Acad Sci USA2008,105:9175-9180.
49. Cooke BD., Rabbit haemorrhagic disease: field epidemiology and the management of wild rabbitpopulations. Rev Sci Tech2002,21:347-358.
50. D’Alession J M&Gerard G F., Second-strand cDNA synthesis with E.coli DNA polylmerase I andRNase H: the fate of information at the mRNA5’ terminus and the effect of E.coli DNA ligase.Nucleic Acid Res.1988,16:1999-2014.
51. Dalpke A.H., Thomssen R., Ritter K., Oxidative injury to endothelial cells due toEpstein-Barrvirus-induced autoantibodies against manganese superoxide dismutase. J Med Virol2003,71:408-416.
52. Delibes-Mateos M., Delibes M., Ferreras P., Villafuerte R: Key role of European rabbits in theconservation of the Western Mediterranean basin hotspot. Conserv Biol2008,22:1106-1117.
53. Dreyfuss G., Matunis M J., Pino-lRoma S., et al. HnRNP proteins and the biogenesis of mRNA.Annu Rev Bi ochem,1993,62(1):289-321
54. Eckerkorn C., et al., Mass spectrometric analysis of blotted proteins after gel electrophoresisseparation by matrix-assisted laser desorption. Electrophoresis13,1992,664-665.
55. Elliott PR., Bilton D., Lomas DA., Lung polymers in Z alpha1-antitrypsin defciency-relatedemphysema. Am J Respir Cell Mol Biol,1998,18(5):670-674
56. Esteves PJ., Abrantes J., Carneiro M., et al., Detection of positive selection in the major capsidprotein VP60of the rabbit haemorrhagic disease virus (RHDV). Virus Res2008,137:253-256.
57. Farkas T., Sestak K., Wei C., et al., Characterization of a rhesus monkey calicivirus representing anew genus of Caliciviridae. J Virol2008,82:5408-5416.
58. Farnos O., Boue O., Parra F., High-level expression and immunogenic properties of therecombinant rabbit hemorrhagic disease virus VP60capsid protein obtained in Pichia pastoris.Journal of Biotechnology.2005,117(3):215-224.
59. Farnos O., Rodriguez D., Valdes O., et al., Molecular and antigenic characterization of rabbithemorrhagic disease virus isolated in Cuba indicates a distinct antigenic subtype. Arch Virol2007,152:1215-1221.
60. Fleming LE., Oquendo S., Bean JA., et al., Pilot detection study of alpha antitrypsin def-ciency ina targeted population. Am J Med Genet,2001,103(1):69-74
61. Gibb JA., Williams JM., The rabbit in New Zealand. In The European rabbit: the history andbiology of a successful colonizer. Edited by Corbet GB, Flux JEC, Rogers PM, Arthur CP, SoriguerRC, Myers K, Parer I, Wood D, Cooke BD, Gibb JA Williams JM, Fenner F, Ris J, Thompson HV,King CM. Oxford: Oxford University Press;1994:158-200.
62. Gildea TR., Shermock KM., Singer ME., et al., Cost-effectiveness analysis of augmentationtherapy for severe al-pha1-antitrypsin defciency. Am J Respir Crit Care Med,2003,167(10):1387–1392
63. Golemis EA., Brent R., Fused moietles inhibit DNA binding by LexA derivatives. Mol Cell Biol,1992,(12):3006-3014.
64. Granzow H., Weiland F., Strebelow HG., et al., Rabbit hemorrhagic disease virus (RHDV):ultrastructure and biochemical studies of typical and core-like particles present in liverhomogenates Virus Res,1996,41:163-172.
65. Gregg DA., House C., Meyer R., et al., Viral haemorrhagic disease of rabbits in Mexico:epidemiology and viral characterization. Rev Sci Tech1991,10:435-451..
66. Harrington P.R., Lindesmith L., Yount B., et al. Binding of Norwalk virus-like particles to ABHhisto-bloodgroup antigens is blocked by antisera from infected human volunteers or experimentallyvaccinated mice. J. Virol.2002,76,12335-12343
67. Henzel W.J., Billeci T.M., et al., Identifying proteins from two-dimensional gels by molecular masssearching of peptide fragments in protein sequence databases. Proc. Natl Acad. Sci.USA90,5011-5015(1993).
68. Hofstetter H., Schambock A., VanDenBerg J., et al., Specific expression of the inserted DNAsegment from hybrid plasmids constructed by the poly(dA).poly(dT)method. Biochin Biophys Acta1976,454(3):587-591.
69. Hunt SA, Baker DW, Chin MH, et al., ACC/AHA guidelines for the evaluation and management ofchronic heart failure in the adult: executive summary. A report of the ACC/AHA task force onpractice guideline.J Am Collcardio,2001,35(2):101-2113.
70. Hutson A.M., Atmar R.L., Marcus D.M., et al., Norwalkvirus-like particle hemagglutination bybinding to h histo-blood group antigens. J. Virol.2003,77,405-415.
71. Issaq H.J., Veenstra T.D., Conrads T.P., et al., The SELDI-TOF MS approach to proteomics: proteinprofiling and biomarker identification. Biochemical and biophysical research communications2002,292:587-592.
72. Jensen P. K., et al., Probing proteomes using capillary isoelectric focusing-electrosprayionizationFourier transform ion cyclotron resonance mass spectrometry. Anal. Chem.1999,71,2076-2084.
73. Juan Barcena, Nuria Verdaguer, Ramon Roca, et al., The coat protein of Rabbit hemorrhagicdisease virus contains a molecular switch at the N-terminal region facing the inner surface of thecapsid. Virology.2004,322(1):118-134.
74. Kawai T., Takahashi K., Sato S., et al., IPS-1, an adaptor triggering RIG-I-and Mda5-mediated typeI interferon induction. Nat Immunol.2005,6(10):981-8.
75. Kiledjian M., and Dreyfuss G., Primary structure and binding activity of the hnRNP U protein:binding RNA through RGG box.1992, EMBO J.11,2655-2664.
76. Larsson C., Natural history and life expectancy in severealpha1-antitrypsin defciency. Pi Z. ActaMed Scand,1978,204(5):345-351
77. Lavazza A., Capucci L., How Many Caliciviruses are there in Rabbits? A Review on RHDV andCorrelated Viruses Lagomorph Biology. Edited by Alves PC, Ferrand N, Hackl nder K: SpringerBerlin Heidelberg;2008:263-278.
78. Lavazza A., Scicluna MT., Capucci L., Susceptibility of hares and rabbits to the European brownhare syndrome virus (EBHSV) and rabbit haemorrhagic disease virus (RHDV) under experimentalconditions. Zentralbl Veterinarmed B1996,43:401-410.
79. Le Gall-Recule G., Zwingelstein F., Laurent S., et al., Phylogenetic analysis of rabbit haemorrhagicdisease virus in France between1993and2000, and the characterisation of RHDV antigenicvariants. Arch Virol2003,148:65-81.
80. Le Gall-ReculéG., Zwingelstein F., Laurent S., et al., Phylogenetic analysis of rabbit haemorrhagicdisease virus in France between1993and2000, and the characterisation of RHDV antigenicvariants. Arch Virol2003,148:65-81.
81. L'Homme Y., Sansregret R., Plante-Fortier E., et al., Genomic characterization of swinecaliciviruses representing a new genus of Caliciviridae. Virus Genes2009,39:66-75.
82. Li, J., et al., Integration of microfabricated devices to capillary electrophoresis—electrospraymassspectrometry using a low dead volume connection: application to rapid analyses ofproteolyticdigests. Anal. Chem.1999,71,3036-3045.
83. Liu SJ, Xue HP, Pu BQ, et al., A new viral disease in rabbit. Anim Husb Vet Med1984,16:253-255.
84. Lowe GDO., Rumley A., Whincup PH., et al., Hemostatic and rheological variables and risk ofcardiovascular disease. Semin Vasc Med,2002,2:429-440.
85. Mariani TD., Halluin K., et al., The Production and Analysis of Genetically Engineered MaleSterile Plants of Maize. In: Abstr35th Ann Maize Genetics Conf. Fargo：University Press，1993,35:40-45.
86. Martinez-Torrecuadrada JL., Cortes E., Vela C., et al., Antigenic structure of the capsid protein ofrabbit haemorrhagic disease virus. J Gen Virol1998,79:1901-1909.
87. McDonald, T., Sheng, S., Stanley, B., et al., Expanding the subproteome of the inner mitochondriausing protein separation technologies: one-and two-dimensional liquid chromatography andtwo-dimensional gel electrophoresis. Mol Cell Proteomics5,2392-2411.
88. Mehdaoui S., Touze A., Laurent S., et a1., Gene transfer using recombinant rabbit hemorrhagicdisease virus capsids with genetically modified DNA encapsidation capacity by addition ofpackaging sequences from or L2protein of human papillomavirus type16. J Virol.2000,74:10332-10340.
89. Meyers G., Wirblich C., Thiel HJ., Rabbit hemorrhagic disease virus--molecular cloning andnucleotide sequencing of a calicivirus genome. Virology1991,184:664-676
90. Mitro S, Krauss H: Rabbit hemorrhagic disease: a review with special reference to its epizootiology.Eur J Epidemiol1993,9:70-78.
91. Neil J D., Nucleotide sequence of the capsid protein gene of two serotypes of San Miguel sea lionvirus: identification of conserved and non-conserved amino acid sequences among caliciviruscapsid proteins. Virus Res.1992,24(2):211-222.
92. Nuwaysir, L.&Stults, J. T., ESI mass spectrometry of phosphopeptides isolated byon-lineimmobilized metal affinity chromatography. J. Am. Soc. Mass Spectrom.1993,4,662–669.
93. O'Farrell P.H., High resolution two-dimensional electrophoresis of proteins. J Biol Chem1975,250:4007-4021
94. Ohlinger VF., Haas B., Meyers G., et al., Identification and characterization of the virus causingrabbit hemorrhagic disease. J Virol1990,64:3331-3336.
95. Okayama H.&Berg P., High efficiency cloning of full-length cDNA. Mol.Cell.Bilo.1982(2):161-170.
96. P, Devaux and D, Gerlier, Antibody cross-reactivity with CD46and lack of cell surface expressionsuggest that moesin might not mediate measles virus binding. J Virol1997,71:1679~1682.
97. Park NY., Chong CY., Kim JH., et al., An outbreak of viral haemorrhagic pneumonia (tentativename) of rabbits in Korea. J Korean Vet Med Assoc1987,23:603-610.
98. Parra F., Prieto M., Purification and characterization of a calicivirus as the causative agent of alethal hemorrhagic disease in rabbits. J Virol1990,64:4013-4015.
99. Pastorino B., Boucomont-Chapeaublanc E., Peyrefitte C.N., et al., Identification of cellularproteome modifications in response to West Nilevirus infection. Mol Cell Proteomics2009,8:1623-1637.
100. Paulo JA., Lee LS., Wu B., et al., Optimized sample preparation of endoscopic collected pancreaticfluid for SDS-PAGE analysis. Elec-trophoresis.2010;31(14):2377-87.
101. Prasad BV., Rothnagel R., Jiang X., et al., Three-dimensional structure of baculovirus-expressedNorwalk virus capsids. J Virol1994,68:5117-5125.
102. Pugnale P., Latorre P., Rossi C., et al., Real-time multiplex PCR assay to quantify hepatitis C virusRNA in peripheral blood mononuclear cells. J Virol Methods,2006,133(2):1952204.
103. Qiyan J., Xifeng J., Fei Y., et al., Short cyclic peptides derived from the C-terminal sequenceof a1-antitrypsin exhibit signifcant anti-HIV-1activity. Bioorganic&Medicinal Chemistry Letters,2012,22:2393–2395.
104. Reichel C., Sudhop T., Braun B., et al., Elevated soluble tumour necrosis factor receptor serumconcentrations and short-term mortality in liver cirrhosis without acute infections.Digestion,2000,62(1):44-51．
105. Rockey DC., Noninvasive assessment of liver fibrosis and portal hypertension with transientelastography．Gastroenterology,2008,134(1):8-14．
106. Rodak L., Smid B., Valicek L., et al., Enzyme-linked immunosorbent assay of antibodies to rabbithaemorrhagic disease virus and determination of its major structural proteins. J Gen Virol1990,71:1075-1080.
107. Roman I., Figys J., Steurs G., et al., In vitro interactions between the two mitochondrial membraneproteins VDAC and cytochrome coxidase. Biochemistry2005,44(39):13192-13201.
108. Roulston A., Marcellus R.C., Branton P.E., Viruses and apoptosis. Annu Rev Microbiol1999,53:577-628
109. Schirrmeier H., Reimann I., Kollner B., et al., Pathogenic, antigenic and molecular properties ofrabbit haemorrhagic disease virus (RHDV) isolated from vaccinated rabbits: detection andcharacterization of antigenic variants. Arch.Virol.1999,144:719-735.
110. Takahashi S., Reddy SV., Chirgwin JM., et al., Cloning and identification of annexin II as anautocrine/Paracrine factor that increases osteoclast formation and bone resorption. J Biol Chem.1994,269(46):296-702.
111. Thouvenin E., Laurent S., Madelaine MF., et al., Bivalent binding of a neutralising antibody to acalicivirus involves the torsional flexibility of the antibody hinge. J Mol Biol1997,270:238-246
112. Tong, A., Wu, L., Lin, Q., et al. Proteomic analysis of cellular protein alterations using a hepatitis Bvirus-producing cellular model. Proteomics,2008,2012-2023.
113. Torrecuadradan M J L., Cortes E., Vela C., et al., Antigenic structure of the capsid protein of rabbithemorrhagic disease virus. J Gen Virol.1998,79(Pt8):1901-1909.
114. Tsui N B., Ngek., Loym., Molecular analysis of circulating RNA in plasma. Methods Mol Biol,2006,336:123-134.
115. Ushijima H., Miyamura.T, Takeda N., Proteolytic processing of sapovirus ORF1polyprotein. JVirol.2005,79:7283-7290.
116. Valicek L., Smid B., Rodah L., et al., Electron and immunoelectron microscopy of rabbithaemorrhagic disease virus (RHDV). Arch Virol,1990,112(3-4):271-275.
117. Van Dinten L C., Johan A., Wassenaar A L M., et al., An infectious arterivirus cDNA clone:Identification of a replicase point mutation that abolishes discontinuous mRNA transcription. ProcNatl Acad Sci. USA.1997,94:991-996.
118. Vester D., Rapp E., Kluge S., et al., Virus-host cell interactions in vaccineproduction cell linesinfected with different human influenza A virus variants: a proteomic approach. Journal ofproteomics2010,73:1656-1669.
119. Wiese S., Reidegeld K.A., Meyer H.E., et al., Protein labeling by iTRAQ: a new tool forquantitative mass spectrometry in proteome research. Proteomics2007,7:340-350
120. Wirblich C., Sibilia M., Boniotti MB., et al.,3C-like protease of rabbit hemorrhagic disease virus:identification of cleavage sites in the ORF1polyprotein and analysis of cleavage specificity. JVirol.1995,69(11):7159-7168.
121. Xu WY., Viral haemorrhagic disease of rabbits in the People's Republic of China: epidemiologyand virus characterization. Rev Sci Tech1991,10:393-408.
122. Y. Doi., M. Kurita., M. Matsumoto., et al., Moesin is not a receptor for Measles Virus entry intomouse embryonic stem cells. J.Virol.1998,72(2):1586-1592.
123. Yasmin MY., Aziz B., et al., Prealbumin rather than albumin is a more sensitive indicator of acuteliver disease.Malays J Pathol,1993,15(2):147.
124. Zhao Z., Huang X., Li N., et al., Direct cloning of cell differential expression genes withfull-length by a new strategy based on the multiple rounds of long distance polymerase chainreaction and magnetic beads mediated subtraction. J Bio technol1999,73:35-41.
125. Zheng X., Hong L., Shi L., et al., Proteomics analysis of host cells infected with infectious bursaldisease virus. Mol Cell Proteomics7,2008,612-6.