Abstract
The classical baculovirus display system (BDS) has often recruited fields including gene delivery, gene therapy, and the genetic engineering of vaccines, as it is capable of presenting foreign polypeptides on the membranes of recombinant baculovirus through a transmembrane protein. However, classical BDS’s high cost, complicated operation, low display efficiency and its inability to simultaneously display multiple gene products impede its practicality. In this study, we present a novel and highly efficient display system based on ires-dependent gp64 for rescuing gp64-null Bacmid of baculovirus construction without affecting the viral replication cycle, which we name the baculovirus multigene display system (BMDS). Laser scanning confocal microscopy demonstrated that eGFP, eYFP, and mCherry were translocated on the membrane of Spodoptera frugiperda 9 cell successfully as expected. Western blot analysis further confirmed the presence of the fluorescent proteins on the budded, mature viral particles. The results showed the display efficiency of target gene on cell surface is fourfold that of classical BDS. In addition, a recombinant baculovirus displaying three kinds of fluorescent proteins simultaneously was constructed, thereby demonstrating the effectiveness of BMDS as a co-display system.
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Acknowledgements
We would like to thank Jonathan Jih (University of California, Los Angeles) for revising the grammar of the manuscript. This study was funded by the National Natural Science Foundation of China (Grant Number 31372373, 31372381), the Natural Science Foundation of Guangdong Province, China (Grant Number 2016A030311018) and Science and Technology Planning Project of Guangzhou, China (Grant Number 201510010276).
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HZ, LY, and JS coordinated the project. HZ and XW performed the research. HZ and JS wrote the manuscript. LY and JS contributed new methods and improved the manuscript. FR, SZ, LX, and MF performed the data analysis. HZ, LY, and JS interpreted the context of results. All authors have read and approved the manuscript.
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Communicated by S. Hohmann.
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Zheng, H., Wang, X., Ren, F. et al. Construction of a highly efficient display system for baculovirus and its application on multigene co-display. Mol Genet Genomics 293, 1265–1277 (2018). https://doi.org/10.1007/s00438-018-1459-9
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DOI: https://doi.org/10.1007/s00438-018-1459-9