| 犬细小病毒HL-01株的分离鉴定及VP2基因真核表达质粒的构建 |
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| 引用本文： | 王玉玲,范京惠,边亚娟,张炳丽,唐丽杰,李一经.犬细小病毒HL-01株的分离鉴定及VP2基因真核表达质粒的构建[J].东北农业大学学报,2006,37(1):69-73. |
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| 作者姓名： | 王玉玲 范京惠 边亚娟 张炳丽 唐丽杰 李一经 |
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| 作者单位： | 东北农业大学动物医学院,黑龙江,哈尔滨,150030 |
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| 摘 要： | 从临床发病犬采集粪便样品,以F81传代细胞进行病毒分离,经血球凝集实验(HA)和血球凝集抑制实验(HI)初步鉴定为犬细小病毒。为进一步确诊,根据Genbank中已发表的犬细小病毒VP2基因序列设计并合成一对引物,通过聚合酶链反应(PCR)扩增出CPV VP2基因,酶切,测序加以鉴定。其序列与国际已发表的CPV-d(type 2)、CPV-1(5 type 2a)、CPV-3(9 type 2b)VP2序列同源性分别为98.97%,98.75%,98.69%,氨基酸序列的同源性分别为98.12%,97.60%,97.60%,从而证明此分离株为犬细小病毒。在获得VP2基因的基础上,为实现VP2蛋白的表达,构建了真核表达质粒pMel BacC-VP2。
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| 关 键 词： | 犬细小病 VP2基因 克隆 表达质粒 构建 |
| 文章编号： | 1005-9369（2006）01-0069-05 |
| 收稿时间： | 2004-11-24 |
| 修稿时间： | 2004年11月24 |
Identification of HL-1 isolate of canine parvovirus and construction of eukaryotic expression plasmid of VP2 gene |
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| WANG Yu-ling,FAN Jing-hui,BIAN Ya-juan,ZHANG Bing-li,TANG Li-jie,LI Yi-jing.Identification of HL-1 isolate of canine parvovirus and construction of eukaryotic expression plasmid of VP2 gene[J].Journal of Northeast Agricultural University,2006,37(1):69-73. |
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| Authors: | WANG Yu-ling FAN Jing-hui BIAN Ya-juan ZHANG Bing-li TANG Li-jie LI Yi-jing |
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| Institution: | College of Veterinary Medicine, Northeast Agrieuhural University, Harbin Heilongjiang 150030, PRC |
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| Abstract: | The HL-01 isolate of canine parvovirus was propagated on F81 cell.And identified by the haem agglutination test(HA) and haem agglutination inhibition test(HI).Using a pair of primers based on the published sequence of CPV′s VP2 gene,the full length gene encoding outer capsid protein VP2 was amplified from cell culture by polymerase chain reaction(PCR).The VP2 gene of HL-01 isolate of canine parvovirus was inserted into pMD18-T vector and was identified by sequencing,digestion.The result showed that there was a high homology innucleotide sequence and aminoacid sequence incomparison with CPV-d(type 2),CPV-15(type 2a) and CPV-39(type 2b).The rate of nucleotide sequence homology was 98.97%,98.75%,98.69%,and the aminoacid homology was 98.12%,97.60%,97.60%.The VP2 gene of the isolate was inserted into the eukaryotic expression plasmid pMel BacC and construced recombinated plasmid pMel BacC-VP2 in order to realize the expression of VP2 protein. |
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| Keywords: | canine parvovirus VP2 cloning pMel BacC expression plasmid construction |
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