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Potato Virus M-Like Nanoparticles: Construction and Characterization

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Abstract

Virus-like particles (VLPs) are multisubunit self-assembly competent protein structures with identical or highly related overall structure to their corresponding native viruses. To construct a new filamentous VLP carrier, the coat protein (CP) gene from potato virus M (PVM) was amplified from infected potato plants, cloned, and expressed in Escherichia coli cells. As demonstrated by electron microscopy analysis, the PVM CP self-assembles into filamentous PVM-like particles, which are mostly 100–300 nm in length. Adding short Gly-Ser peptide at the C-terminus of the PVM, CP formed short VLPs, whereas peptide and protein A Z-domain fusions at the CP N-terminus retained its ability to form typical PVM VLPs. The PVM-derived VLP carrier accommodates up to 78 amino acid-long foreign sequences on its surface and can be produced in technologically significant amounts. PVM-like particles are stable at physiological conditions and also, apparently do not become disassembled in high salt and high pH solutions as well as in the presence of EDTA or reducing agents. Despite partial proteolytic processing of doubled Z-domain fused to PVM VLPs, the rabbit IgGs specifically bind to the particles, which demonstrates the functional activity and surface location of the Z-domain in the PVM VLP structure. Therefore, PVM VLPs may be recognized as powerful structural blocks for new human-made nanomaterials.

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Acknowledgments

The authors wish to thank Prof. Dr. P. Pumpens for critical reading of the manuscript and suggestions, Prof. Dr. K. Tars and Dr. A. Kazaks for helpful discussions. G. Grinberga, G. Resevica, and V. Zeltina are acknowledged for their technical assistance. This work was supported by the ERAF Grant 2013/0052/2DP/2.1.1.1.0/13/APIA/VIAA/019.

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  1. Latvian Biomedical Research and Study Centre, Ratsupites 1, Riga, 1067, Latvia

    Ieva Kalnciema, Ina Balke, Dace Skrastina, Velta Ose & Andris Zeltins

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  1. Ieva Kalnciema
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Correspondence to Andris Zeltins.

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Kalnciema, I., Balke, I., Skrastina, D. et al. Potato Virus M-Like Nanoparticles: Construction and Characterization. Mol Biotechnol 57, 982–992 (2015). https://doi.org/10.1007/s12033-015-9891-0

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