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Citation: Ning XIONG, Yong ZHANG, Yao WANG, Bao-yu YANG, Shi-yun CHEN. Isolation and Identification of Canine Parvovirus Serotype 2a and Its VP2 Protein Expression in Transgenic Tobacco [J].VIROLOGICA SINICA, 2008, 23(3) : 203-210.  http://dx.doi.org/10.1007/s12250-008-2928-9

Isolation and Identification of Canine Parvovirus Serotype 2a and Its VP2 Protein Expression in Transgenic Tobacco

  • Corresponding author: Shi-yun CHEN, sychen@wh.iov.cn
  • Received Date: 13 December 2007
    Accepted Date: 10 April 2008
    Available online: 01 June 2008

    Fund Project: National Science Foundation of China 30771239

  • A strain of canine parvovirus(CPV)was isolated from feces of an ill puppy in an animal hospital in Wuhan, China. It was designated as CPV/WH02/06. This isolate was identified as serotype CPV-2a by the hemagglutination test, CPV Ag detection strip, electron microscopy, and PCR. The vp2 gene was cloned and sequenced and assigned GenBank accession number EU377537. A 1242 bp segment of the 5’ region of the vp2 gene was cloned and inserted into the binary vector pBI121 and used for Agrobacterium-mediated tobacco transformation. Transgenic tobacco plants were selected on MS medium supplemented with 100 μg/mL kanamycin and 100 μg/mL timentin. Integration of the vp2 gene into the tobacco genome was confirmed by PCR using T1 progeny plants, and the expression of the VP2 protein was confirmed by Western blotting.

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    Isolation and Identification of Canine Parvovirus Serotype 2a and Its VP2 Protein Expression in Transgenic Tobacco

      Corresponding author: Shi-yun CHEN, sychen@wh.iov.cn
    • 1. State Key Laboratory of Virology, Wuhan Institute of Virology, the Chinese Academy of Sciences, Wuhan 430071, China
    • 2. Graduate School of the Chinese Academy of Sciences, Beijing 100049, China
    Fund Project:  National Science Foundation of China 30771239

    Abstract: A strain of canine parvovirus(CPV)was isolated from feces of an ill puppy in an animal hospital in Wuhan, China. It was designated as CPV/WH02/06. This isolate was identified as serotype CPV-2a by the hemagglutination test, CPV Ag detection strip, electron microscopy, and PCR. The vp2 gene was cloned and sequenced and assigned GenBank accession number EU377537. A 1242 bp segment of the 5’ region of the vp2 gene was cloned and inserted into the binary vector pBI121 and used for Agrobacterium-mediated tobacco transformation. Transgenic tobacco plants were selected on MS medium supplemented with 100 μg/mL kanamycin and 100 μg/mL timentin. Integration of the vp2 gene into the tobacco genome was confirmed by PCR using T1 progeny plants, and the expression of the VP2 protein was confirmed by Western blotting.