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.

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, 
  • 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.

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    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.

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