Purification of Tomato Bushy Stunt Virus Particles by One-Step Hydroxyapatite Column Chromatography

Authors

  • Zh. Tleukulova L.N. Gumilyov Eurasian University, 2 Satpayev Str., Nur-Sultan, Kazakhstan
  • Z. Stamgaliyeva L.N. Gumilyov Eurasian University, 2 Satpayev Str., Nur-Sultan, Kazakhstan
  • A. Dildabek School of Sciences and Humanities, Nazarbayev University, 53 Kabanbay Batyr Ave., Nur-Sultan, Kazakhstan
  • G. Mukiyanova L.N. Gumilyov Eurasian University, 2 Satpayev Str., Nur-Sultan, Kazakhstan
  • R.T. Omarov L.N. Gumilyov Eurasian University, 2 Satpayev Str., Nur-Sultan, Kazakhstan

DOI:

https://doi.org/10.18321/ectj1131

Keywords:

virus particles, Tomato bushy stunt virus, purification, hydroxyapatite column chromatography

Abstract

The main aim of this work was to develop a time-saving and cost-effective purification method of infectious plant viral nanoparticles. Virions of Tomato bushy stunt virus (TBSV), which is a member of Tombusvirus genus, were purified by one-step Bio-gel HT Hydroxyapatite (HA) column chromatography. Extracts from Nicotiana benthamiana plants infected with TBSV were directly loaded onto the HA column and eluted by 10 mM sodium phosphate buffer (pH 6.8). A specificity of virions has been confirmed by immunoblotting and electron microscopy. Homogeneity of virions was tested by SDS-PAGE, where only 41 kDa polypeptide bands referring to the capsid protein of TBSV were detected by Coomassie staining. The biological infectious activity of a purified material was demonstrated by observing TBSV-specific symptoms observed in N. benthamiana plants at 7‒10 days of post-inoculation (dpi). Moreover, purified virions were used for immunization of the BALb/c mouse to raise primary antibodies against the TBSV virus. Our results show that in low concentrations of sodium phosphate buffer total proteins extracted from infected plants adsorb to HA sorbent, while viral particles do not adsorb to the HA matrix and flow throw column due to Ca2+ ions implicated in TBSV virions’ structure. This highly effective and simple virus purification protocol can also be used for the isolation of other plant virions.

References

(1). K.-B.G. Scholthof, S. Adkins, H. Czosnek, P. Palukaitis, E. Jacquot, T. Hohn, B. Hohn, K. Saunders, T. Candresse, P. Ahlquist, C. Hemenway, G.D. Foster, Mol. Plant Pathol. 12 (2011) 938–954. Crossref DOI: https://doi.org/10.1111/j.1364-3703.2011.00752.x

(2). S.C. Harrison, A.J. Olson, C.E. Schutt, F.K. Winkler, G. Bricogne, Nature 276 (1978) 368– 373. Crossref DOI: https://doi.org/10.1038/276368a0

(3). P. Hopper, S.C. Harrison, R.T. Sauer, J. Mol. Biol. 177(1984) 701–713. Crossref DOI: https://doi.org/10.1016/0022-2836(84)90045-7

(4). A.J. Olson, G. Bricogne, S.C. Harrison, J. Mol. Biol. 171 (1983) 61–93. Crossref DOI: https://doi.org/10.1016/S0022-2836(83)80314-3

(5). P.Q. Hearne, D.A. Knorr, B.I. Hillman, T.J. Morris, Virology 177 (1990) 141–151. Crossref DOI: https://doi.org/10.1016/0042-6822(90)90468-7

(6). A. Llauró, E. Coppari, F. Imperatori, A.R. Bizzarri, J.R. Caston, L. Santi, S. Cannistraro, P.J. de Pablo, Biophys. J. 109 (2015) 390–397. Crossref DOI: https://doi.org/10.1016/j.bpj.2015.05.039

(7). H. Peyret, J. Virol. Methods 225 (2015) 59–63. Crossref DOI: https://doi.org/10.1016/j.jviromet.2015.09.005

(8). E. Schröder, T. Jönsson, L. Poole, Anal. Biochem. 313 (2003) 176–178. Crossref DOI: https://doi.org/10.1016/S0003-2697(02)00567-5

(9). L.J. Cummings, M.A. Snyder, K. Brisack, Method Enzymol. 463 (2009) 387–404. Crossref DOI: https://doi.org/10.1016/S0076-6879(09)63024-X

(10). M. Niimi, T. Masuda, K. Kaihatsu, N. Kato, S. Nakamura, T. Nakaya, F. Arai, Sensor. Actuat. B-Chem. 201 (2014) 185–190. Crossref DOI: https://doi.org/10.1016/j.snb.2014.04.011

(11). G.G. Markov, I.G. Ivanov, Anal. Biochem. 59 (1974) 555–563. Crossref DOI: https://doi.org/10.1016/0003-2697(74)90309-1

(12). G. Bernardi, M.-G. Giro, C. Gaillard, Biochimica et Biophysica Acta (BBA) - Protein Structure 278 (1972) 409–420. Crossref DOI: https://doi.org/10.1016/0005-2795(72)90001-3

(13). G.M. Kostner, A. Holasek, Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism 488 (1977) 417–431. Crossref DOI: https://doi.org/10.1016/0005-2760(77)90200-4

(14). G.P. Smith, T.R. Gingrich, BioTechniques 39 (2018). Crossref DOI: https://doi.org/10.2144/000112032

(15). PM 7/126 (1) Electron microscopy in diagnosis of plant viruses. EPPO Bulletin 45 (2015) 450– 453. Crossref DOI: https://doi.org/10.1111/epp.12260

(16). T.M. Yergaliyev, Zh. Nurbekova, G. Mukiyanova, A. Akbassova, M. Sutula, S. Zhangazin, A. Bari, Zh. Tleukulova, M. Shamekova, Zh.K. Masalimov, R.T. Omarov, Plant Physiol. Biochem. 109 (2016) 36–44. Crossref DOI: https://doi.org/10.1016/j.plaphy.2016.09.001

(17). A.B. Dildabek, Z.B. Stamgaliyeva, B.B. Ilyasova, Z.B. Tleukulova, A.A. Madirov, S.M. Kassenova, R.T. Omarov, S.K. Naekova, A.Zh. Akbassova, Russ. J. Plant Physiol. 68 (2021) 883–889. Crossref DOI: https://doi.org/10.1134/S102144372104004X

Downloads

Published

31-12-2021

How to Cite

Tleukulova, Z., Stamgaliyeva, Z., Dildabek, A., Mukiyanova, G., & Omarov, R. (2021). Purification of Tomato Bushy Stunt Virus Particles by One-Step Hydroxyapatite Column Chromatography. Eurasian Chemico-Technological Journal, 23(4), 277‒282. https://doi.org/10.18321/ectj1131

Issue

Vol. 23 No. 4 (2021)

Section

Articles

License

You are free to: Share — copy and redistribute the material in any medium or format. Adapt — remix, transform, and build upon the material for any purpose, even commercially.

Eurasian Chemico-Technological Journal applies a Creative Commons Attribution 4.0 International License to articles and other works we publish.

Subject to the acceptance of the Article for publication in the Eurasian Chemico-Technological Journal, the Author(s) agrees to grant Eurasian Chemico-Technological Journal permission to publish the unpublished and original Article and all associated supplemental material under the Creative Commons Attribution 4.0 International license (CC BY 4.0).

Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI.