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| Co-expression of Ebola Virus GP and VP40 Proteins and Virus-like particles Assembly in Mammalian Cells |
| SUN Peng-yan1,2,3,4, LI Kui1,2,3,4, LIU Cun-bao2,3,4, YAO Yu-feng2,3,4, CHU Xiao-jie2,3,4, BAI Hong-mei2,3,4, YANG Xu2,3,4, HUANG Wei-wei2,3,4, SUN Wen-jia2,3,4, MA Yan-bing2,3,4 |
1. Kunming Medical University, Kunming 650500, China;
2. Institute of Medical Biology, Chinese Academy of Medical Science & Peking Union Medical College, Kunming 650118, China;
3. Yunnan Key Laboratory of Research and Development on Severe Infection Disease, Kunming 650118, China;
4. Yunnan Engineering Research Center of Vaccine Research and Development on Severe Infection Disease, Kunming 650118, China |
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Abstract Objective: To co-expression Ebola virus proteins GP and VP40 in mammalian cells and generate Ebola virus-like particles. Methods: According to amino acid sequences, the nucleotide sequences of Ebola-Zaire GP and VP40 genes were optimized based on the codon usage bias in mammalian cells. The synthesized genes were cloned into expression plasmids pcDNA3.1 or pBudCE4.1 which has two expression units, and then 293FT cells were transfected with the recombinant plasmids using lipofectamine2000. The expression of recombinant proteins was detected by Western blot, and the VLPs were observed by electron microscope. Results: Specific reactive bands recognized by anti-GP antibody were found in cells transfected with the recombinant plasmid carrying optimized GP gene. The expression of recombinant GP protein mediated by the plasmid pBudCE4.1/GP/VP40 which co-expressing GP and VP40 was significantly higher than that mediated by co-transfection with the plasmids pcDNA3.1/GP which expresses GP and pcDNA3.1/VP40 which expresses VP40. Besides, VP40 expression was detectable although the level was low. Classical Ebola virus-like particles were found under the observation with electron microscope. Conclusion: Ebola virus proteins GP and VP40 were successfully expressed and assembled into virus-like particles in 293FT cells, which laid an important foundation for further studies on the development of a vaccine or virus detection reagent.
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Received: 21 June 2016
Published: 25 December 2016
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Cite this article:
SUN Peng-yan, LI Kui, LIU Cun-bao, YAO Yu-feng, CHU Xiao-jie, BAI Hong-mei, YANG Xu, HUANG Wei-wei, SUN Wen-jia, MA Yan-bing. Co-expression of Ebola Virus GP and VP40 Proteins and Virus-like particles Assembly in Mammalian Cells. China Biotechnology, 2016, 36(12): 66-71.
URL:
https://manu60.magtech.com.cn/biotech/DOI:10.13523/j.cb.20161210 OR https://manu60.magtech.com.cn/biotech/Y2016/V36/I12/66
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