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Comparison and opti...
Comparison and optimization of protein extraction and two-dimensional gel electrophoresis protocols for liverworts
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- Yadav, Sandhya (författare)
- Banaras Hindu Univ, Ctr Adv Study Bot, Inst Sci, Dept Bot, Varanasi 221005, Uttar Pradesh, India.
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- Srivastava, Akanksha (författare)
- Banaras Hindu Univ, Ctr Adv Study Bot, Inst Sci, Dept Bot, Varanasi 221005, Uttar Pradesh, India.
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- Biswas, Subhankar (författare)
- Banaras Hindu Univ, Ctr Adv Study Bot, Inst Sci, Dept Bot, Varanasi 221005, Uttar Pradesh, India.
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- Chaurasia, Neha (författare)
- North Eastern Hill Univ, Dept Biotechnol & Bioinformat, Shillong 793022, Meghalaya, India.
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- Singh, Sushil Kumar (författare)
- Bot Survey India Northern Reg Ctr, 192 Kaulagarh Rd, Dehra Dun 248003, Uttarakhand, India.
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- Kumar, Sanjiv (författare)
- KTH,Glykovetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova
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- Srivastava, Vaibhav (författare)
- KTH,Glykovetenskap,Albanova VinnExcellence Center for Protein Technology, ProNova
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- Mishra, Yogesh (författare)
- Banaras Hindu Univ, Ctr Adv Study Bot, Inst Sci, Dept Bot, Varanasi 221005, Uttar Pradesh, India.
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Banaras Hindu Univ, Ctr Adv Study Bot, Inst Sci, Dept Bot, Varanasi 221005, Uttar Pradesh, India North Eastern Hill Univ, Dept Biotechnol & Bioinformat, Shillong 793022, Meghalaya, India. (creator_code:org_t)
- 2020-02-07
- 2020
- Engelska.
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Ingår i: BMC Research Notes. - : Springer Nature. - 1756-0500. ; 13:1
- Relaterad länk:
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https://doi.org/10.1...
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https://bmcresnotes....
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Objective Liverworts possess historical adaptive strategies for abiotic stresses because they were the first plants that shifted from water to land. Proteomics is a state-of-the-art technique that can capture snapshots of events occurring at the protein level in many organisms. Herein, we highlight the comparison and optimization of an effective protein extraction and precipitation protocol for two-dimensional gel electrophoresis (2-DE) of liverworts. Results We compared three different protein extraction methods, i.e.,1.5 M Tris-HCl (pH 8.8), 50 mM Tris-HCl (pH 7.5), and polyvinylpolypyrrolidone (PVPP) extraction, followed by three precipitation methods, i.e., 80% ethanol, 80% acetone, and 20% tricholoroacetic acid (TCA)-acetone, in a liverwort Dumortiera hirsuta. Among these methods, 50 mM Tris-HCl (pH 7.5) extraction, followed by 20% TCA-acetone precipitation, appeared to be more suitable for 2-DE. Furthermore, we performed modifications during protein washing, re-solubilization in rehydration buffer and isoelectric focusing (IEF). The modifications provided us better results in terms of protein yield, resolution, spot numbers, and intensities for 2-DE gels of D. hirsuta and other two liverworts, i.e., Marchantia paleacea and Plagiochasma appendiculatum. Furthermore, we randomly selected spots from the 2-DE gel of D. hirsuta and identified using mass spectrometry, which confirms the applicability of this protocol for liverworts proteomics.
Ämnesord
- TEKNIK OCH TEKNOLOGIER -- Annan teknik -- Livsmedelsteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Other Engineering and Technologies -- Food Engineering (hsv//eng)
Nyckelord
- Dumortiera hirsuta
- Liverworts
- Marchantia paleacea
- Plagiochasma appendiculatum
- Proteomics
- Two-dimensional gel electrophoresis (2-DE)
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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