Journal of Jilin University Medicine Edition ›› 2013, Vol. 39 ›› Issue (2): 204-208.doi: 10.7694/jldxyxb20130203

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Construction and expression of eukaryotic expression vector of interferon β1a gene

WANG Yan1,2,HUANG Zhi-li1,LI Yan-hui1,ZHANG Li-jun1,XIE Gui-qiu3   

  1. (1.School of Applied Chemistry and Biological Technology,Shenzhen Polytechnic,Shenzhen 518055,China;2.Key Laboratory for Molecular Enzymology and Engineering,Ministry of Education,Collegeof Life Science,Jilin University,Changchun 130012,China;3.Department of Gene Engineering,School of Pharmaceutical Sciences,Jilin University,Changchun 130021,China)
  • Received:2012-09-12 Online:2013-03-28 Published:2013-03-28

Abstract: To construct the CHO expression system of interferon-β1a (IFN-β1a) gene and to explore the expression efficacy of human IFN-β1a gene in eukaryotic cells. Methods The synthesized IFN-β1a gene was amplified and cloned into plasmid pSV2-dhfr.Cysteine 17 was mutated to serine.The constructed recombinant plasmid pSV2-dhfr-IFN-β1a was transfected to CHO-dhfr- cells,and the monoclonal cell strains growing stably were obtained by MTX pressure screening.The genomic DNA of transfected cells was extracted;target gene was identified by PCR.The antiviral activity of IFN-β1a in transfected cells was determined by Wish cytopathic inhibition test.Results  Both PCR and restriction enzyme analysis showed that the recombinant plasmid pSV2-dhfr-IFN-β1a was constructed correctly.The IFN-β1a gene was amplified by using the genomic DNA of transfected cells as template.The expressed IFN-β1a showed high antiviral activity up to 3×105 IU·mL-1.Conclusion A eukaryotic expression vector of IFN-β1a gene is successfully constructed,and the IFN-β1a protein with biological activity is expressed in CHO cells.

Key words: interferon &beta, 1a, eukaryotic cells, CHO cells

CLC Number: 

  • Q784