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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (02): 369-373.doi: 10.3724/SP.J.1006.2011.00369

• RESEARCH NOTES • Previous Articles     Next Articles

Optimization of Extraction Method and Two-Dimensional Electrophoresis Conditions for Proteome Analysis of Jute Functional Leaf

CHEN Fu-Cheng,QI Jian-Min*,XU Jian-Tang,CHEN Tao,TAO Ai-Fen,LIN Pei-Qing,CHEN Mei-Xia,GUO Ying,LI Hua-Li   

  1. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2010-07-06 Revised:2010-10-03 Online:2011-02-12 Published:2010-12-15

Abstract: The protein extraction methods and electrophoresis conditions are the key facts of two-dimensional electrophoresis in proteomics. In this study, in order to set up a suitable method for jute leaf total protein extraction and the optimized condition for two-dimensional electrophoresis, we compared the protein extraction methods using TCA acetone, Tris-base acetone and Tris-HCl. We also optimized the sample loading amount and sample preparation approach. The results showed that protein productivity reached 79.0 mg g–1 at most by TCA acetone method, and 44.2% and 114.1% higher than by other two methods, respectively. Besides, the 2-DE image background was the clearest and the protein spots were the most reaching about 602 by TCA acetone method, which is much more suitable as extraction method for jute leaf protein two-dimensional electrophoresis. The most suitable sample loading quantity was 350 mg, which lays a foundation for further study of jute proteomics. The construction of two-dimensional electrophoresis system can be use to study the differences of protein expression on normal material and mutant of jute, and promote the application of proteomics in jute mutant.

Key words: Jute leaf, Extraction method, Protein two-dimensional electrophoresis (2-DE), Optimizing of loading amount

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