人博卡病毒基因克隆及衣壳编码基因序列变异分析Genome Cloning and Phylogenetic Analysis of Human Bocavirus Capsid Gene
漆正宇;瞿小旺;刘文培;谢志平;高寒春;郑丽舒;匡治州;郁建平;段招军;
摘要(Abstract)：
为了解人博卡病毒(Human Bocavirus,HBoV)VP1基因进化关系;阐明HBoV目前具体的变化规律,用PCR的方法扩增了1株HBoV的全基因和9株HBoV的VP1基因,克隆并测序,在此基础上,将HBoV的全基因序列和衣壳序列分别与细小病毒亚科其他14个有代表性的病毒进行遗传分析,构建进化树,对目前所有可得到的HBoV的17个衣壳蛋白进行二级结构分析和抗原性分析。结果显示:HBoV全基因序列与B19关系较远,但衣壳序列遗传关系较近。以有典型性的猫瘟细小病毒(Feline parvovirus,FPV)衣壳蛋白为参照,分析多个HBoV衣壳序列之间的变异情况,显示HBoV衣壳的二级结构基础表现出较高的保守性,序列之间的变化主要发生在高抗原区域和感染活性区域。衣壳病毒变异情况显示HBoV在稳定自身的情况下表现出一定的活跃性以逃避免疫反应,也表现出一定的感染适应力。
关键词(KeyWords)： 人博卡病毒;克隆;序列分析;B19;猫瘟细小病毒
基金项目(Foundation): 国家自然科学基金资助项目(30600682)
作者(Authors): 漆正宇;瞿小旺;刘文培;谢志平;高寒春;郑丽舒;匡治州;郁建平;段招军;
参考文献(References)：
- [1]Simon A,Groneck P,Kupfer B,et al.Detection of bo-cavirus DNA in nasopharyngeal aspirates of a child withbronchiolitis[J].J Infect,2007,in press,Available fromhttp://www.sciencedirect.com
- [2]Allander T,Tammi M T,Eriksson M,et al.Cloning ofa human parvovirus by molecular screening of respiratorytract samples[J].Proc Natl Acad Sci USA,2005,102:12891-12896.
- [3]侯云德.急性呼吸道病毒感染的病原学和防治[M].北京:中国协和医科大学出版社,2005:6-10.
- [4]瞿小旺,漆正宇,段招军,等.儿童急性呼吸道博卡病毒感染[J].病毒学报,2006,22(2):79-82.
- [5]Qu X W,Duan Z J,Qi Z Y,et al.Human Bocavirus In-fection,People′s Republic of China[J].Emerg InfectDis,2007,13(1):165-168.
- [6]段招军,黄灿平,瞿小旺,等.中国内陆发现HCoV-HKU1感染及N和S蛋白基因序列及进化分析[J]。病毒学报,2006,22(4):241-247.
- [7]赵林清,钱渊,朱汝南,等.北京地区婴幼儿急性呼吸道感染与新近报道的人细小病毒相关性的初步研究[J].中华微生物学和免疫学杂志,2006,26(5):385-388.
- [8]曾爱平,林峰,杨宁敏,等.荧光定量PCR方法检测儿童下呼吸道感染的人博卡病毒[J].浙江检验医学,2006,4(4):34-36.
- [9]Agbandje M,McKenna R,Rossmann M G,et al.Struc-ture determination of feline panleukopenia virus emptyparticles[J].Proteins,1993,16:155-171.
- [10]Cotmore S F,Tattersall P:Characterization and molec-ular cloning of a human parvovirus genome[J].Science1984,226:1161-1165.
- [11]程凌鹏,陈森雄,Zhou Z H,等.伊蚊浓核病毒三维结构的比较分析[J].中国科学C辑:生命科学,2006,36(4):355-359.
- [12]Agbandje-McKenna M,Rossmann M G.The Structureof Human Parvovirus B19[M].//Anderson L J,YoungN S(eds).Human Parvovirus B19.Monogr Virol,1997,20:3-15.
- [13]Kajigaya S,Fujii H,Field A,et al.Self-assembled B19parvovirus capsids,produced in a baculovirus system,are antigenically and immunogenically similar to nativevirions[J].Proc Natl Acad Sci USA,1991,88:4646-4650.
- [14]Brown K E,Anderson S M,Young N S.Erythrocyte Pantigen:Cellular receptor for B19 parvovirus[J].Sci-ence,1993,262:114-117.
- [15]Zoltan Za dori,Szelei 1 J,Marie-Claude L,et al.A vi-ral phospholipase A2 is required for parvovirus infectivi-ty[J].Develop Cell,2001,1:291-302.
- [16]赵林清,钱渊,朱汝南,等.北京地区新近报道的人Boca病毒VP1、VP2蛋白编码区基因序列分析[J].中华微生物学和免疫学杂志,2006,26,(12):1087-1091.
- [17]Agbandje M,McKenna R,Rossmann M G,et al.Structure determination of feline panleukopenia virusempty particles[J].Proteins,1993,16:155-171.
- [18]Strassheim M L,Gruenberg A,Veijalainen P,et al.Two dominant neutralizing antigen determinants of ca-nine parvovirus are found on the threefold spike of thevirus capsid[J].Virology,1994,198:175-184.
- [19]Wikoff W R,Wang G,Parrish C R,et al.The struc-ture of a neutralized virus:Canine parvovirus complexedwith neutralizing antibody fragment[J].Structure,1994,2:595-607.
- [20]Langeveld J P M,Casal J I,Vela C,et al.B-cellepitopes of canine parvovirus:Distribution of the prima-ry structure and exposure on the viral surface[J].J Vir-ol,1993,67:765-772.
- [21]Chapman M S,Rossmann M G.Structure,sequenceand function correlations among Parvoviruses[J].Vi-rology,1993,194:491-508.
- [22]Schierup M H,Hein.Recombination and the molecularclock[J].Mol Biol Evol,2000,17:1578-1579.
- [23]Huelsenbeck J P,Rannala,Phylogenetic methods comeof age:Testing hypotheses in a phylogenetic context[J].Science,1997,276:227-232.
- [24]Schierup M H,Hein.Consequences of recombinationon traditional phylogenetic analysis[J].Genetics,2000,156:879-891.
- [25]Shackelton L A,Parrish C R,et al.High rate of viralevolution associated with the emergence of carnivoreparvovirus[J].Proc Natl Acad Sci USA,2005,102:379-384.
- [26]Shackelton L A,Holmes E C.Phylogenetic evidencefor the rapid evolution of human B19 erythrovirus[J].JVirol,2006,80(7):3666-3669.