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Expression and purification of classical swine fever virus E2 protein from Sf9 cells using a modified vector

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Abstract

Objective

To develop a simple method for efficient expression of classical swine fever virus (CSFV) E2 protein.

Results

The pFastBac HT B vector (pFastHTB-M1) was modified by adding a melittin signal peptide sequence. The E2 gene fragment without the transmembrane region was cloned into pFastHTB-M1. The modified vector has clear advantage over the original one, as evidenced by the purified recombinant E2 protein that was detected significantly by SDS-PAGE.

Conclusions

The modified vector has the potential for large-scale production and easy purification of the CSFV E2 protein or other proteins of interests.

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Acknowledgements

This work is funded by the National Youth Science Fund of NSFC (National Natural Science Foundation of China) (31101793) and the Ministry of Education (20120101130014).

Supporting information

Supplementary Fig. 1—Schematic map of pFastHTB-M1 vector construction from pFastBacTM HT B (a) and sequencing confirmation (b).

Supplementary Fig. 2—Identification of the recombinant baculovirus genome containing the E2 gene fragment of classical swine fever virus using universal M13 primers and E2-specific primers.

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Authors and Affiliations

  1. Institute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310058, China

    Ling Yang, Xingmeng Lu & Weihuan Fang

Authors
  1. Ling Yang
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  2. Xingmeng Lu
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  3. Weihuan Fang
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Corresponding author

Correspondence to Weihuan Fang.

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Yang, L., Lu, X. & Fang, W. Expression and purification of classical swine fever virus E2 protein from Sf9 cells using a modified vector. Biotechnol Lett 39, 1821–1825 (2017). https://doi.org/10.1007/s10529-017-2426-y

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  • DOI: https://doi.org/10.1007/s10529-017-2426-y

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