Protein extraction from Phoenix dactylifera L. leaves, a recalcitrant material, for two-dimensional electrophoresis

被引：38

作者：

Gomez-Vidal, Sonia ^{[1
]}

Tena, Manuel ^{[2
]}

Lopez-Llorca, Luis Vicente ^{[1
]}

Salinas, Jesus ^{[1
]}

机构：

[1] Univ Alicante, Multidisciplinary Inst Environm Studies Ramon Mar, Dept Marine Sci & Appl Biol, Plant Pathol Lab, Alicante 03080, Spain

[2] Univ Cordoba, ETSIAM, Dept Biochem & Mol Biol, Cordoba, Spain

来源：

ELECTROPHORESIS | 2008年 / 29卷 / 02期

关键词：

2-DE; leaf protein extraction; Phoenix dactylifera;

D O I：

10.1002/elps.200700380

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

This work was aimed at optimizing a protein extraction procedure for date palm (Phoenix dactylifera L.) leaves, a highly recalcitrant plant tissue for 2-DE. Five protein extraction protocols based on different protein precipitation agents (TCA/acetone vs. phenol (Ph) methods) and protein resolubilization methods (physical treatments, e.g., sonication, shaking and/or heating) were tested. Ph/SDS extraction with methanol/ammonium acetate precipitation, followed by DOC preincubation and TCA/acetone precipitation and, finally, solubilization by shaking in rehydration solution was found to be the best protein extraction method. We conclude that DOC with TCA/acetone precipitation step eliminates interfering compounds, thus allowing efficient resolubilization of date palm leaf proteins. This method could be appropriate for proteomic studies such as date palm colonization by entomo-pathogenic fungi.

引用

页码：448 / 456

页数：9

共 41 条

[31] A simple protocol for protein extraction of recalcitrant fruit tissues suitable for 2-DE and MS analysis [J].

Song, Jun ;

Braun, Gordon ;

Bevis, Eric ;

Doncaster, Kristen .

ELECTROPHORESIS, 2006, 27 (15) :3144-3151

[32]

THOMAS TC, 1990, METHOD ENZYMOL, V182, P499

[33] Efficient extraction of proteins from woody plant samples for two-dimensional electrophoresis [J].

Valcu, Cristina-Maria ;

Schlink, Katja .

PROTEOMICS, 2006, 6 (14) :4166-4175

[34] Optimization of protein extraction and solubilization for mature grape berry clusters [J].

Vincent, Delphine ;

Wheatley, Matthew D. ;

Cramer, Grant R. .

ELECTROPHORESIS, 2006, 27 (09) :1853-1865

[35] SOLUBILIZATION OF LOW-DENSITY LIPOPROTEIN WITH SODIUM DEOXYCHOLATE AND RECOMBINATION OF APOPROTEIN-B WITH DIMYRISTOYLPHOSPHATIDYLCHOLINE [J].

WALSH, MT ;

ATKINSON, D .

BIOCHEMISTRY, 1983, 22 (13) :3170-3178

[36] Protein extraction for two-dimensional electrophoresis from olive leaf, a plant tissue containing high levels of interfering compounds [J].

Wang, W ;

Scali, M ;

Vignani, R ;

Spadafora, A ;

Sensi, E ;

Mazzuca, S ;

Cresti, M .

ELECTROPHORESIS, 2003, 24 (14) :2369-2375

[37] A universal and rapid protocol for protein extraction from recalcitrant plant tissues for proteomic analysis [J].

Wang, Wei ;

Vignani, Rita ;

Scali, Monica ;

Cresti, Mauro .

ELECTROPHORESIS, 2006, 27 (13) :2782-2786

[38]

WESTERMEIER R., 2002, Proteomics in Practice A Laboratory Manual

[39] An efficient protein preparation for proteomic analysis of developing cotton fibers by 2-DE [J].

Yao, Yuan ;

Yang, Yi-Wei ;

Liu, Jin-Yuan .

ELECTROPHORESIS, 2006, 27 (22) :4559-4569

[40] Phenolic compounds in date palm cultivars sensitive and resistant to Fusarium oxysporum [J].

Ziouti, A ;

ElModafar, C ;

Fleuriet, A ;

ElBoustani, S ;

Macheix, JJ .

BIOLOGIA PLANTARUM, 1996, 38 (03) :451-457

← 1 2 3 4 5 →