| 带有组氨酸标签的马传染性贫血病毒感染性分子克隆的构建 |
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| 引用本文： | 温建新,仇华吉,涂亚斌,王柳,彭金美,童光志.带有组氨酸标签的马传染性贫血病毒感染性分子克隆的构建[J].中国预防兽医学报,2005,27(3):193-195. |
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| 作者姓名： | 温建新 仇华吉 涂亚斌 王柳 彭金美 童光志 |
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| 作者单位： | 中国农业科学院,哈尔滨兽医研究所,兽医生物技术国家重点实验室,黑龙江,哈尔滨,150001 |
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| 基金项目： | 国家高技术研究发展计划(863计划)，国家自然科学基金 |
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| 摘 要： | 马传染性贫血(EIA)弱毒疫苗的广泛使用存在野毒和疫苗毒鉴别困难的问题.本研究以已构建的马传染性贫血驴白细胞弱毒疫苗株的感染性分子克隆(pOK8266)为基础,在其S2基因内引入NspV酶切位点,将人工合成的编码6个组氨酸的寡核苷酸插入NspV位点,获得带有组氨酸标签的重组质粒pOK8266-HIS.将pOK8266-HIS转染驴白细胞,将驴白细胞转染产物传至第6代时,在电镜下观察到了典型的马传染性贫血病毒粒子.提取pOK8266-HIS衍生病毒的前病毒基因组DNA,通过PCR扩增和测序表明,衍生病毒基因组中引入了6个组氨酸标签,从而获得了带有分子标志的马传染性贫血弱毒疫苗株,为野毒株和疫苗病毒的鉴别诊断奠定了基础.本研究还证明了S2基因中的插入突变并不影响马传染性贫血病毒的体外复制.
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| 关 键 词： | 马传染性贫血病毒 感染性分子克隆 组氨酸标签 |
| 文章编号： | 1008-0589(2005)03-0193-03 |
| 修稿时间： | 2004年4月23日 |
Generation of a histidine-tagged equine infectious anemia virus |
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| WEN Jian-xin,QIU Hua-ji,TU Ya-bin,WANG Liu,PENG Jin-mei,TONG Guang-zhi.Generation of a histidine-tagged equine infectious anemia virus[J].Chinese Journal of Preventive Veterinary Medicine,2005,27(3):193-195. |
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| Authors: | WEN Jian-xin QIU Hua-ji TU Ya-bin WANG Liu PENG Jin-mei TONG Guang-zhi |
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| Abstract: | Equine infectious anemia virus (EIAV) donkey leukocyte attenuated vaccine has been applied extensively in China,but a method being able to differentiate wild_type EIAV strains from the vaccine strain is not available.In this study,an NspV site was introduced into the S2 gene of pOK8266, an infectious molecular clone derived from EIAV vaccine virus,and then a 6_histidine tag was introduced into NspV site,generating pOK8266_HIS.The pOK8266_HIS was transfected into donkey leukocytes.Cytopathogenic effects were found in donkey leukocytes infected with the virus rescued from pOK8266_HIS after six passages.EIAV virions were observed by electron microscope.Sequence analysis indicated that the proviral DNA of the rescued virus contains the histidine tag.The results also demonstrated that insertion of short sequence in S2 gene does not affect in vitro replication of EIAV in target cells. |
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| Keywords: | equine infectious anemia virus infectious molecular clone His-tag marker |
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