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EIAV感染性分子克隆、嵌合病毒构建及生物学特性研究

批准号30170706 学科分类石油、天然气地质学 ( D0207 )
项目负责人杨香华 负责人职称 依托单位中国农业科学院哈尔滨兽医研究所
资助金额0.00
万元
项目类别面上项目 研究期限2002 年 01 月 01 日 至
2004 年 12 月 01 日
中文主题词马传染性贫血病毒,感染性分子克隆,嵌合病毒
英文主题词equine infectious anemia virus (EIAV), infectious molecular clone, chimeric virus

摘要

中文摘要 克隆EIAV弱毒及其亲本强毒L株各基因片段,顺次连接,获得强弱毒全长前病毒克隆并测序鞠赴秆〕鲇懈腥拘缘娜醵局昵安《痉肿涌寺。溆隠株序列差异大的区域用L株翁婊唬竦们咳跚逗喜《荆萌醵靖腥拘苑肿涌寺⊙苌牟《炯扒逗喜《窘又纸】德恚觳馄渖镅匦浴6匝芯咳醵局氯跫懊庖呋坪妥魑滩《锬P途哂兄匾庖濉
英文摘要 The complete genomes of donkey leukocyte attenuated equine infectious anemia virus (DLA-EIAV) and EIAV strain Liaoning (L-EIAV) proviruses were cloned and sequenced.The infectious molecular clone (pOK8266)of DLA-EIAV was constructed and its derived virus(vOK8266) was obtained. After comparing the gemome of DLA-EIAV with that of L-EIAV,three chimeric molecular clone were constructed based on pOK8266 by replaced the long terminal repeat (LTR) sequence or/and envelope gene of DLA-EIAV with the counterpart of L-EIAV.A chimeric virus(vOKVltr) with LTR substitution was generated.Horses vaccinated with these two cloned viruses were subjected to determination of the clinic symptom and pathological lesion,detection of antibody response and virus load in the plasma.The results indicated that,though it's LTR was substituted,the virulence of vOKVltr was not stronger than it's parental virus ,but it's replication competence increased ,which resulted in the better immunogenicity.In addition,we found some variation in the LTR and env gene which may associate with the immunity and pathogenesis of DLA-EIAV.These results will help to further study of the molecular mechanism of attenuation and protection of DLA-EIAV.
结题摘要 本研究以马传贫驴白细胞弱毒疫苗(DLA-EIAV)感染细胞总DNA及其亲本强毒EIAV L株感染马外周血白细胞中DNA为模板,应用PCR方法分段扩增出DLA-EIAV和L株的前病毒DNA,并分别获得EIAV弱毒和强毒的全长DNA克隆。转染驴白细胞后获得了EIAV驴白细胞弱毒的感染性分子克隆及其衍生病毒。比较强毒和弱毒的基因组序列,将其中变异较大的区域(LTR和env)进行了替换,获得了3个强、弱毒嵌合克隆。其中以弱毒感染性分子克隆为骨架,构建的用强毒LTR替换了弱毒LTR的嵌合克隆获得了衍生病毒。将这些克隆毒接种马,通过临床及病理组织学观察、各结构蛋白抗体反应监测以及血液中病毒定量检测,结果发现用强毒的LTR置换弱毒的LTR并没有使弱毒株毒力增强,但在体内的复制能力有所增强,因而其免疫原性好于弱毒感染性分子克隆株,说明仅改变LTR不会影响马传贫病毒的毒力。通过对试验马体内不同时期LTR和env基因进行监测,发现了一些可能与免疫保护和致病性有关的变异,这为进一步研究马传贫病毒驴白细胞弱毒的致弱和免疫保护机制奠定了基础。

成果

序号 标题 类型 作者
1 马传染性贫血病强弱毒基因组序列差异及其生物学意义 会议 童光志|
2 马传染性贫血强/弱毒嵌合病毒的体外构建 会议 涂亚斌|王柳|仇华吉|温建新|谷守林|陈焕春|童光志|
3 Proviral Genomic Sequence Analysis of Chinese Donkey Leukocyte Attenuated Equine Infectious Anemia Virus Vaccine and its Parental Virus Strain Liaoning 期刊 L.Wang|G.Z.Tong|H.Q.Liu|Z.B.Yang|H.J.Qiu|X.G.Kong|M.Wang|
4 Complete sequence of proviral DNA of equine infectious anemia virus strain L 期刊 Liu HQ|Wang L|Yang ZB|Kong XG|and Tong GZ|

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