Serving the Society Since 1986
Advanced Search
Volume 19 Issue 2
April 2004
    Citation: WANG Hai-zhen, Yang Song, SU Xiao-yun, CAO Rui-bing, CHEN Pu-yan. Construction and Expression of the Prokaryotic Expressing Vector Containing Tandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene .VIROLOGICA SINICA, 2004, 19(2) : 171-173.
    shu

    Construction and Expression of the Prokaryotic Expressing Vector Containing Tandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene

    • 1.

      1. Key Lab of Animal Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China

    • 2.

      College of Animal Science and Veterinary Medicine of Shenyang Agricultural University, Shenyang 110161,China

    • Corresponding author: CHEN Pu-yan, 
    • Available online: 20 April 2004
    • Full E2 gene fragment of Classical swine fever virus (CSFV)shimen strain was amplified by RT-PCR method, then a specific fraction -the A, D antigenic domains- , which encodes protein with good antigenicity and suitable for being expressed in E. coli with high level, was amplified respectively by 2 different pair of primers. Then the 2 amplified fragments were tandemly linked and inserted into prokaryotic expressing vector pET-32a to obtain the plasmid pET-2e. The SDS-PAGE assay showed that although the proteins were present in the form of inclusion body, the linked proteins were expressed from plasmid pET-2e as expected, and can be expressed with high level when induced with IPTG.Western-blotting showed good antigenicity of the target protein.
    • 加载中
    • 加载中
    PDF

    Article Metrics

    Article views(3932) PDF downloads(965) Cited by()

    Related
    Proportional views

      Construction and Expression of the Prokaryotic Expressing Vector Containing Tandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene

        Corresponding author: CHEN Pu-yan,
      • 1. 1. Key Lab of Animal Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
      • 2. College of Animal Science and Veterinary Medicine of Shenyang Agricultural University, Shenyang 110161,China

      Abstract: Full E2 gene fragment of Classical swine fever virus (CSFV)shimen strain was amplified by RT-PCR method, then a specific fraction -the A, D antigenic domains- , which encodes protein with good antigenicity and suitable for being expressed in E. coli with high level, was amplified respectively by 2 different pair of primers. Then the 2 amplified fragments were tandemly linked and inserted into prokaryotic expressing vector pET-32a to obtain the plasmid pET-2e. The SDS-PAGE assay showed that although the proteins were present in the form of inclusion body, the linked proteins were expressed from plasmid pET-2e as expected, and can be expressed with high level when induced with IPTG.Western-blotting showed good antigenicity of the target protein.

        HTML

      Relative (20)

      目录

      This journal is a member of, and subscribes to the principles of, the Committee on Publication Ethics (COPE)

      鄂ICP备05001977号

         

      鄂公网安备 42010602003674号

      Copyright © 2019 Virologica Sinica. All rights reserved

      /

      DownLoad:  Full-Size Img  PowerPoint
      Return
      Return