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Izvorni znanstveni članak
https://doi.org/10.5562/cca1804
Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues
Dubravko Pavoković
orcid.org/0000-0001-7198-6063
; Department of Molecular Biology, Division of Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, HR-10000 Zagreb, Croatia
Bojana Križnik
; Department of Molecular Biology, Division of Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, HR-10000 Zagreb, Croatia
Marijana Krsnik-Rasol
; Department of Molecular Biology, Division of Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, HR-10000 Zagreb, Croatia
| Puni tekst: engleski, pdf (2 MB) | str. 177-183 | citiraj | |
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APA 6th Edition Pavoković, D., Križnik, B. i Krsnik-Rasol, M. (2012). Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues. Croatica Chemica Acta, 85 (2), 177-183. https://doi.org/10.5562/cca1804 MLA 8th Edition Pavoković, Dubravko, et al. "Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues." Croatica Chemica Acta, vol. 85, br. 2, 2012, str. 177-183. https://doi.org/10.5562/cca1804. Citirano 07.03.2021. Chicago 17th Edition Pavoković, Dubravko, Bojana Križnik i Marijana Krsnik-Rasol. "Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues." Croatica Chemica Acta 85, br. 2 (2012): 177-183. https://doi.org/10.5562/cca1804 Harvard Pavoković, D., Križnik, B., i Krsnik-Rasol, M. (2012). 'Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues', Croatica Chemica Acta, 85(2), str. 177-183. https://doi.org/10.5562/cca1804 Vancouver Pavoković D, Križnik B, Krsnik-Rasol M. Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues. Croatica Chemica Acta [Internet]. 2012 [pristupljeno 07.03.2021.];85(2):177-183. https://doi.org/10.5562/cca1804 IEEE D. Pavoković, B. Križnik i M. Krsnik-Rasol, "Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues", Croatica Chemica Acta, vol.85, br. 2, str. 177-183, 2012. [Online]. https://doi.org/10.5562/cca1804 |
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Sažetak
Plant tissues contain relatively low amounts of proteins whose extraction is often difficult due to the presence of interfering compounds such as rigid cellulosic cell wall, storage polysaccharides, lipids and other contaminants that can cause protein degradation or modification. Therefore it is important to optimize protein extraction and to establish a robust protocol for two-dimensional gel electrophoresis (2-DE) and downstream processing. In this study, acetone, trichloroacetic acid/acetone and Tris-buffered phenol/methanol extraction protocols were evaluated on non-model and recalcitrant plants: a Beta vulgaris L. in vitro cell line, Mammillaria gracilis Pfeiff. in vitro plants and Sempervivum tectorum L. leaves. Although phenol extraction was more time-consuming than the other two methods, it gave almost two-fold higher protein yield, and spectral analysis showed less contamination. SDS-PAGE, 2-DE and bioinformatic analysis showed that protein extraction using phenol was superior to the other two methods, providing more protein bands or spots on the gels and less proteolysis. These results lead to the conclusion that the phenol method is the most suitable protein extraction method for these non-model and recalcitrant plant tissues. (doi: 10.5562/cca1804)
Ključne riječi
electrophoresis; mass spectrometry; phenol; plant proteomics; protein extraction; trichloroacetic acid
Hrčak ID: 80970
URI
https://hrcak.srce.hr/80970
Moj profil
Srce