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Volume 21 Issue 4
August 2006
    Citation: HUANG Xin, GAO Ya-ping, WANG Xi-liang, XIN Zhong-tao, DONG Jie, SHAO Ning-sheng, LIU Chuan. Screening and Identification of the SARS-CoV N Protein Epitopes .VIROLOGICA SINICA, 2006, 21(4) : 314-318.
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    Screening and Identification of the SARS-CoV N Protein Epitopes

    • 1.

      1.Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China

    • 2.

      Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China

    • Available online: 20 July 2006
    • The epitopes of SARS-CoV were screened from a 12-mer phage display random peptide library using anti- SARS-CoV horse polyclonal antibodies of as a target. The phage clones enriched in biopannings were sequenced. Two consensus sequences were obtained, DXXDP and TXTLL, which had close identity to the 341-345 aa and 392-396 aa of SARA-CoV N protein sequences,respectively. The phage clones with consensus sequences and the N protein were both recognized and bound by the antibodies in a competitive-inhibition ELISA test. The consensus sequence peptides were cloned and displayed on the bacterial flagellin. Balb/c mice were vaccinated using the reconstruted bacteria and the collected serum was shown to speccifically combined with SARS-CoV N protein. This confirmed that DXXDP and TXTLL are two continuous epitopes of SARS-CoV N protein.
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      Screening and Identification of the SARS-CoV N Protein Epitopes

      • 1. 1.Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China
      • 2. Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China

      Abstract: The epitopes of SARS-CoV were screened from a 12-mer phage display random peptide library using anti- SARS-CoV horse polyclonal antibodies of as a target. The phage clones enriched in biopannings were sequenced. Two consensus sequences were obtained, DXXDP and TXTLL, which had close identity to the 341-345 aa and 392-396 aa of SARA-CoV N protein sequences,respectively. The phage clones with consensus sequences and the N protein were both recognized and bound by the antibodies in a competitive-inhibition ELISA test. The consensus sequence peptides were cloned and displayed on the bacterial flagellin. Balb/c mice were vaccinated using the reconstruted bacteria and the collected serum was shown to speccifically combined with SARS-CoV N protein. This confirmed that DXXDP and TXTLL are two continuous epitopes of SARS-CoV N protein.

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