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 suitabl...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.展开更多
文摘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.
