ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2005, Vol. 36 ›› Issue (12): 1318-1322.

• 兽医 • Previous Articles     Next Articles

Expression of CSFV E2 Envelope Protein Gene in Eukaryotic Cells and Preparation of Monoclonal Antibodies against CSFV E2 Envelope Protein

XU Xin-gang;HU Jian-he;ZHANG Yan-ming; ZHANG Hai-tang; CHEN Yong-yao   

  1. 1. College of Animal Science and Technology, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100,China; 2. Department of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-12-25 Published:2005-12-25

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Abstract: The recombinant retroviral vector pBABE-puro-E2 was constructed by inserting E2 gene full-length cDNA of CSFV Shimen strain into pBABE-puro. Both the recombinant retroviral vector and pVSVg plasmid were transfected into 293T cells by calcium phosphate transfection method, thus the pseudovirus were produced. The pseudovirus infected eukaryotic cells SP2/0 and expression E2 protein was determined by puromycin-resistant and FACS analysis.BALB/c mice were injected in abdomen with expressing E2 protein SP2/0 cells. Anti-CSFV E2 antibody was screened by FACS. The results showed that CSFV E2 protein was expressed in SP2/0 cells’ envelope protein successfully. FACS could detect specific anti-E2 antibody in immune mouse serum. Mouse spleen cells were fused with myeloma SP2/0. Culture supernatants of hybridoms were screened by FACS and four cell lines which could secrete against E2 protein of CSFV McAbs stably were obtained. The McAbs not only react with CSFV specially but also had neutralization activity. The McAbs type belongs to IgM.

Key words: CSFV, E2 envelope protein, retroviral vector, pseudovirus, FACS, McAbs