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Applied Biochemistry and Biotechnology

, Volume 173, Issue 8, pp 2129–2139 | Cite as

Enhanced Expression of GCRV VP6 in CIK Cells by Relative Sequence Optimization

  • Bo XiaoEmail author
  • Xiaoyan Chi
  • Li Zhang
  • Huige Qu
  • Yujing Liu
  • Xiaojie Wang
  • Juhua Zhou
Article

Abstract

Efficient expression of target protein is one of strategies for gene therapy or vaccine design. Many studies showed that codon optimization could enhance the expression of target proteins. In this paper, a target sequence of about 1.26 kb encoding the major capsid protein VP6 of grass carp reovirus (GCRV) and an optimized counterpart were synthesized and inserted into vectors for expressing VP6. The final constructs (named as pcDV6G and pcDV6YG) were transfected in Ctenopharyngodon idellus kidney (CIK) cells. The fluorescence analysis and the Western blot results showed that the gene fragment was transfected and expressed in CIK cells successfully. Although the qRT-PCR results showed no difference at the messenger RNA (mRNA) levels between the different versions of vp6 in the indicated stages, the enzyme-linked immunosorbent assay (ELISA) results showed that the protein level of VP6 expressed by pcDV6YG was higher than that by pcDV6G in the indicated hours. Taken together, these results suggest that the enhanced expression of GCRV VP6 in CIK cells by relative sequence optimization may be a good choice for making DNA vaccine against GCRV.

Keywords

Grass carp reovirus Capsid protein Recombinant vector Transfection Eukaryotic expression 

Notes

Acknowledgments

This study was supported by the Science and Technology Development Project of Yantai, Shandong, China (2011066), and the talent funds of Ludong University (LY2008337).

Conflict of Interest

There is no conflict of interest to disclose. All authors contributed to the preparation of the manuscript and agreed the version.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Bo Xiao
    • 1
    Email author
  • Xiaoyan Chi
    • 1
  • Li Zhang
    • 1
  • Huige Qu
    • 1
  • Yujing Liu
    • 1
  • Xiaojie Wang
    • 1
  • Juhua Zhou
    • 1
  1. 1.School of Life SciencesLudong UniversityYantaiPeople’s Republic of China

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