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Virologica Sinica, 19 (2) : 171-173, 2004
Brief Reports
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. 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
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.
  Published online: 20 Apr 2004
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