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Volume 25 Issue 2
April 2010
    Citation: Jian WANG, Hong-yan GUO, Rui JIA, Xuan XU, Juan TAN, Yun-qi GENG, Wen-tao QIAO. Preparation of BFV Gag Antiserum and Preliminary Study on Cellular Distribution of BFV* .VIROLOGICA SINICA, 2010, 25(2) : 115-122.  http://dx.doi.org/10.1007/s12250-010-3110-8
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    Preparation of BFV Gag Antiserum and Preliminary Study on Cellular Distribution of BFV*

    cstr: 32224.14.s12250-010-3110-8

    • Key Laboratory of Molecular Microbiology and Biotechnology, Ministry of Education; Key Laboratory Microbial Functional Genomics in Tianjin; College of Life Sciences, Nankai University, Tianjin 300071, China

    • Corresponding author: Wen-tao QIAO, wentaoqiao@nankai.edu.cn.
    • Received Date: 09 November 2009
      Accepted Date: 16 December 2009
      Available online: 01 April 2010

      Fund Project: Grant from State Key Laboratory for Infectious Diseases Prevention and Control, SKL 2008SKLID310The Key Project of the Ministry of Education of China 108028National Natural Science Foundation of China 3090068

    • Viruses (e.g. Human immunodeficiency virus, Human simplex virus and Prototype foamy virus) are obligate intracellular parasites and therefore depend on the cellular machinery for cellular trafficking. Bovine foamy virus (BFV) is a member of the Spumaretrovirinae subfamily of Retroviruses, however, details of its cellular trafficking remain unknown. In this study, we cloned the BFV gag gene into prokaryotic expression vector pET28a and purified the denaturalized Gag protein. The protein was used to immunize BALB/c mouse to produce antiserum, which could specifically recognize the BFV Gag protein in BFV-infected cells through western blot assay. Additionally, these results demonstrated that both the optimal and suboptimal cleavage of Gag protein occur in BFV-infected cells. Subsequently, the Gag antiserum was used to investigate subcellular localization of BFV. In immunofluorescence microscopy assays, colocalization microtubules (MTs) and assembling viral particles were clearly observed, which implied that BFV may transport along cellular MTs in host cells. Furthermore, MTs-depolymerizing assay indicated MTs were required for the efficient replication of BFV. In conclusion, our study suggests that BFV has evolved the mechanism to hijack the cellular cytoskeleton for its replication.
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      Preparation of BFV Gag Antiserum and Preliminary Study on Cellular Distribution of BFV*

        Corresponding author: Wen-tao QIAO, wentaoqiao@nankai.edu.cn.
      • Key Laboratory of Molecular Microbiology and Biotechnology, Ministry of Education; Key Laboratory Microbial Functional Genomics in Tianjin; College of Life Sciences, Nankai University, Tianjin 300071, China
      • Received Date: 09 November 2009
      • Accepted Date: 16 December 2009
      Fund Project:  Grant from State Key Laboratory for Infectious Diseases Prevention and Control, SKL 2008SKLID310The Key Project of the Ministry of Education of China 108028National Natural Science Foundation of China 3090068

      Abstract: Viruses (e.g. Human immunodeficiency virus, Human simplex virus and Prototype foamy virus) are obligate intracellular parasites and therefore depend on the cellular machinery for cellular trafficking. Bovine foamy virus (BFV) is a member of the Spumaretrovirinae subfamily of Retroviruses, however, details of its cellular trafficking remain unknown. In this study, we cloned the BFV gag gene into prokaryotic expression vector pET28a and purified the denaturalized Gag protein. The protein was used to immunize BALB/c mouse to produce antiserum, which could specifically recognize the BFV Gag protein in BFV-infected cells through western blot assay. Additionally, these results demonstrated that both the optimal and suboptimal cleavage of Gag protein occur in BFV-infected cells. Subsequently, the Gag antiserum was used to investigate subcellular localization of BFV. In immunofluorescence microscopy assays, colocalization microtubules (MTs) and assembling viral particles were clearly observed, which implied that BFV may transport along cellular MTs in host cells. Furthermore, MTs-depolymerizing assay indicated MTs were required for the efficient replication of BFV. In conclusion, our study suggests that BFV has evolved the mechanism to hijack the cellular cytoskeleton for its replication.

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