ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2014, Vol. 45 ›› Issue (1): 94-100.doi: 10.11843/j.issn.0366-6964.2014.01.013

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Cloning and Optimizing Expression of E2 Gene of Bovine Viral Diarrhea-mucosal Disease Virus in BCG

ZENG Fan-li1， ZHANG Yun2， ZHANG Meng3， SHI Kun1， LI Jian-ming1， LIU Fei2， LI Jing2， DU Rui4*   

  1. (1.College of Chinese Medicine Materials， Jilin Agricultural University， Changchun 130118， China；2.College of Animal Science &Technology， Jilin Agricultural University， Changchun 130118， China；3.Changchun Kinsey Pharmaceutical Co.Ltd， Changchun 130012， China;4.Key Laboratory of Animal Production，Product Quality and Security of Ministry of Education of the people′s Republic of China， Level 2 Laboratory of Medical Animal of Jilin Province， College of Graduate， Jilin Agricultural University， Changchun 130118， China)
  • Received:2013-07-26 Online:2014-01-23 Published:2014-01-23

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Abstract:

To develop genetic engineering living-vector vaccine of Bovine viral diarrhea-mucosal disease virus (BVDV)， E2 gene of BVDV Changchun184 strain was selected and the complete sequence was analyzed，then the epitopes of E2 protein were predicted by bioinformatics software.Six pairs of primers were designed according to E2 gene sequence，and the epitopes coding regions (1-297，1-345，1-374，45-297，45-345，45-374) of E2 gene were amplified by PCR and cloned into pMV361 vector.PCR，restriction endonuclease digestion and sequence analysis proved that the 6 fragments were inserted into the expected position.It was indicated that the six prokaryotic expression plasmids were constructed.The positive recombinant plasmids were electro-transformed into BCG for expression at 45 ℃.The SDS-PAGE and Western blot results indicated that the recombinant proteins could react with polyclonal antibody against BVDV.Our study provides a basis for the further studies on genetic engineering living-vector vaccine of BVDV.

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