Optimization of Protein Extraction and Two-Dimensional Electrophoresis Protocols for Oil Palm Leaf.
Oil palm (Elaeis guineensis) is an important economic crop cultivated for its nutritional palm oil. A significant amount of effort has been undertaken to understand oil palm growth and physiology at the molecular level, particularly in genomics and transcriptomics. Recently, proteomics studies have begun to garner interest. However, this effort is impeded by technical challenges. Plant sample preparation for proteomics analysis is plagued with technical challenges due to the presence of polysaccharides, secondary metabolites and other interfering compounds. Although protein extraction methods for plant tissues exist, none work universally on all sample types. Therefore, this study aims to compare and optimize different protein extraction protocols for use with two-dimensional gel electrophoresis of young and mature leaves from the oil palm. Four protein extraction methods were evaluated: phenol-guanidine isothiocyanate, trichloroacetic acid-acetone precipitation, sucrose and trichloroacetic acid-acetone-phenol. Of these four protocols, the trichloroacetic acid-acetone-phenol method was found to give the highest resolution and most reproducible gel. The results from this study can be used in sample preparations of oil palm tissue for proteomics work.
Department of Integrative and Applied Biology, Sime Darby Technology Centre Sdn Bhd, 1st Floor, Block B, UPM-MTDC Technology Centre III, Universiti Putra Malaysia, Lebuh Silikon, 43400, Serdang, Selangor, Malaysia, leona.daniela.jefferydaim@simedarby.com.
Leona Daniela Jeffery
LD
Daim
Tony Eng Keong
TE
Ooi
Hirzun Mohd
HM
Yusof
Nazia Abdul
NA
Majid
Saiful Anuar Bin
SA
Karsani
D000096:Acetone; D028023:Arecaceae / Q000737:chemistry*; D005591:Chemical Fractionation / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D019800:Phenol; D018515:Plant Leaves / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D014238:Trichloroacetic Acid
D010940:Plant Proteins; D000096:Acetone; D019800:Phenol; D014238:Trichloroacetic Acid