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A method for protein extraction from different subcellular fractions of laticifer latex in Hevea brasiliensis compatible with 2-DE and MS

BACKGROUND: Proteomic analysis of laticifer latex in Hevea brasiliensis has been received more significant attentions. However, the sticky and viscous characteristic of rubber latex as cytoplasm of laticifer cells and the complication of laticifer latex membrane systems has made it challenge to isol...

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Autores principales: Wang, Xuchu, Shi, Minjing, Lu, Xiuli, Ma, Ruifeng, Wu, Chenggong, Guo, Anping, Peng, Ming, Tian, Weimin
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2898674/
https://www.ncbi.nlm.nih.gov/pubmed/20565811
http://dx.doi.org/10.1186/1477-5956-8-35
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author Wang, Xuchu
Shi, Minjing
Lu, Xiuli
Ma, Ruifeng
Wu, Chenggong
Guo, Anping
Peng, Ming
Tian, Weimin
author_facet Wang, Xuchu
Shi, Minjing
Lu, Xiuli
Ma, Ruifeng
Wu, Chenggong
Guo, Anping
Peng, Ming
Tian, Weimin
author_sort Wang, Xuchu
collection PubMed
description BACKGROUND: Proteomic analysis of laticifer latex in Hevea brasiliensis has been received more significant attentions. However, the sticky and viscous characteristic of rubber latex as cytoplasm of laticifer cells and the complication of laticifer latex membrane systems has made it challenge to isolate high-quality proteins for 2-DE and MS. RESULTS: Based on the reported Borax/PVPP/Phenol (BPP) protocol, we developed an efficient method for protein preparation from different latex subcellular fractions and constructed high-resolution reference 2-DE maps. The obtained proteins from both total latex and C-serum fraction with this protocol generate more than one thousand protein spots and several hundreds of protein spots from rubber particles as well as lutoid fraction and its membranes on the CBB stained 2-DE gels. The identification of 13 representative proteins on 2-DE gels by MALDI TOF/TOF MS/MS suggested that this method is compatible with MS. CONCLUSION: The proteins extracted by this method are compatible with 2-DE and MS. This protein preparation protocol is expected to be used in future comparative proteomic analysis for natural rubber latex.
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spelling pubmed-28986742010-07-08 A method for protein extraction from different subcellular fractions of laticifer latex in Hevea brasiliensis compatible with 2-DE and MS Wang, Xuchu Shi, Minjing Lu, Xiuli Ma, Ruifeng Wu, Chenggong Guo, Anping Peng, Ming Tian, Weimin Proteome Sci Methodology BACKGROUND: Proteomic analysis of laticifer latex in Hevea brasiliensis has been received more significant attentions. However, the sticky and viscous characteristic of rubber latex as cytoplasm of laticifer cells and the complication of laticifer latex membrane systems has made it challenge to isolate high-quality proteins for 2-DE and MS. RESULTS: Based on the reported Borax/PVPP/Phenol (BPP) protocol, we developed an efficient method for protein preparation from different latex subcellular fractions and constructed high-resolution reference 2-DE maps. The obtained proteins from both total latex and C-serum fraction with this protocol generate more than one thousand protein spots and several hundreds of protein spots from rubber particles as well as lutoid fraction and its membranes on the CBB stained 2-DE gels. The identification of 13 representative proteins on 2-DE gels by MALDI TOF/TOF MS/MS suggested that this method is compatible with MS. CONCLUSION: The proteins extracted by this method are compatible with 2-DE and MS. This protein preparation protocol is expected to be used in future comparative proteomic analysis for natural rubber latex. BioMed Central 2010-06-18 /pmc/articles/PMC2898674/ /pubmed/20565811 http://dx.doi.org/10.1186/1477-5956-8-35 Text en Copyright ©2010 Wang et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology
Wang, Xuchu
Shi, Minjing
Lu, Xiuli
Ma, Ruifeng
Wu, Chenggong
Guo, Anping
Peng, Ming
Tian, Weimin
A method for protein extraction from different subcellular fractions of laticifer latex in Hevea brasiliensis compatible with 2-DE and MS
title A method for protein extraction from different subcellular fractions of laticifer latex in Hevea brasiliensis compatible with 2-DE and MS
title_full A method for protein extraction from different subcellular fractions of laticifer latex in Hevea brasiliensis compatible with 2-DE and MS
title_fullStr A method for protein extraction from different subcellular fractions of laticifer latex in Hevea brasiliensis compatible with 2-DE and MS
title_full_unstemmed A method for protein extraction from different subcellular fractions of laticifer latex in Hevea brasiliensis compatible with 2-DE and MS
title_short A method for protein extraction from different subcellular fractions of laticifer latex in Hevea brasiliensis compatible with 2-DE and MS
title_sort method for protein extraction from different subcellular fractions of laticifer latex in hevea brasiliensis compatible with 2-de and ms
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2898674/
https://www.ncbi.nlm.nih.gov/pubmed/20565811
http://dx.doi.org/10.1186/1477-5956-8-35
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