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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (05): 921-927.doi: 10.3724/SP.J.1006.2012.00921

• RESEARCH NOTES • Previous Articles     Next Articles

Optimization of Two-dimensional Electrophoresis Condition for Rice Grain Protein and Comparison of Relevant Proteomic Methods

LI Qi-Song,CHEN Jun,LIN Shi-Sheng,LI Zhong,ZHANG Zhi-Xing,LIN Wen-Xiong*   

  1. Institute of Agricultural Ecology, School of Life Sciences, Fujian Agriculture and Forestry University / Key Laboratory of Crop Physiology and Molecular Ecology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2011-07-08 Revised:2011-12-19 Online:2012-05-12 Published:2012-03-05
  • Contact: 林文雄, E-mail: lwx@fjau.edu.cn E-mail:liqisong0591@163.com

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Abstract: A suitable proteomic strategy for rice grain protein research is important for further understanding rice grain development at proteome level. In this study, three different protein extraction methods were compared and then electrophoretic conditions of hand-made gel lands and IPG lands in IEF (isoelectric focusing) were optimized. Furthermore, three important proteomic research strategies (MALDI-TOF/MS, western-blot and phosphoproteome) were used for identification and analysis of in-gel proteins. The results showed that the method of soluble protein extraction was most suitable for investigation of grain proteomics, with a well-distributed 2-DE gel profile under optimized electrophoretic conditions, and it was confirmed that MALDI-TOF/MS, western-blot and phosphoproteome were the three suitable methods for the analysis of in-gel proteins. The establishment of the suitable system for rice grain proteomic research paves the way for the next step to further analyse rice grain development at proteome level.

Key words: Rice, Grain filling, Proteomics, Two-dimensional electrophoresis, Protein phosphorylation

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