›› 2010, Vol. 37 ›› Issue (3): 65-70.

• 生物技术 • Previous Articles     Next Articles

Cloning，Expression and Immunological Activity Analysis E 2 Gene of Classical Swine Fever Virus Isolates from Sichuan Strain

HAN Guo-quan1,2,GUO Wan-zhu1,2,LIN Hua1,WANG Li-na3, ZHANG Bo1,CHEN Di-shi1,CHEN Yang1   

  1. (1.Animal Biotechnology Center,Sichuan Agricultural University,Ya’an 625014,China; 2.Key Laboratory of Animal Disease and Human Health of Sichuan Province,Sichuan Agricultural University,Ya’an 625014,China; 3.Chengdu Product Quality Supervision and Inspection Institute of Sichuan Province,Chengdu 610041,China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-03-20 Published:2010-03-20
  • Contact: GUO Wan-zhu

Abstract:

In order to investigate the cloning,expression and immunological activity analysis E2 gene of classical swine fever virus (CSFV) isolates from Sichuan. The CSFV total RNA of Sichuan strain was extracted from infected PK-15 cells. The objective E 2 gene was cloned by RT-PCR and directionally cloned into prokaryotic expression vector. The expressing plasmids were transformed into E.coli Rosetta-gami-TM(DE3) plysS. The recombinant bacteria were induced expression by IPTG. The expressed proteins were analyzed by SDS-PAGE and Western blotting. The results indicated that the objective E 2 gene was obtained and contains E2 protein complete codon DNA sequence (CDS), which have 1119 bp，encoding 373 amino acid. The recombinant expression plasmids were successfully constructed of complete CDS and N terminal major antigen gene of CSFV E2 protein,which named pET-E2(pe) and pET-mE2(pe). The recombinant bacteria could express the target fusion proteins which are about 45.32 ku and 28.49 ku by SDS-PAGE detection. The expression of E2 (pe),mE2 (pe) fusion proteins could be recognized by CSFV-positive sera,and had good immunological reaction activity. The research obtained the CSFV E2 gene of Sichuan strain. The E2 (pe)，mE2 (pe) fusion proteins were expressed and have biological activity.

Key words: classical swine fever virus; Sichuan strain; E 2 gene; prokaryotic expression; immunological activity

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