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Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (4): 612-619.doi: 10.3724/SP.J.1006.2010.00612

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Construction of Protein Two-Dimensional Polyacrylamide Gel Electrophoresis System for Brassica napus

GAN Lu1,2,LI Dian-Rong1,3,ZANG Xin4,FU Chun-Hua1,2,YU Long-Jiang1,2,LI Mao-Teng12*   

  1. 1 Institute of Resource Biology and Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 43007, China; 2 Key Laboratory of Molecular Biophysics, Ministry of Education, Wuhan 430074, China; 3 Hybrid Rapeseed Research Center of Shaanxi Province, Dali 715105, China; 4 Bioengineering Department of Zhengzhou University, Zhengzhou 450001, China
  • Received:2009-09-08 Revised:2009-12-24 Online:2010-04-12 Published:2010-02-09
  • Contact: LI Mao-Teng, E-mail: limaoteng426@163.com; limaoteng426@mail.hust.edu.cn

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Abstract:

Two-dimensional polyacrylamide gel electrophoresis has been widely applied in proteomic researches. The approach can analyze dynamic changes of proteomics of biological samples under different conditions to identify specific proteins and genes. The two-dimensional gel systems for rice, Arabidopsis and other species have been established. The purpose of this research was to establish a suitable system for B. napus to analyze proteomics. Rapeseed variety of 08127 was used in the experiment, the proteins were obtained by the trichloroacetic extraction method with pH 4–7 gel brands under the improved IEF procedure, showing a clean two-dimensional electrophoresis map, so a proper system for B. napus wasformed. Then the system was validated by using the young seedlings, leaves and stems from other varieties, and the same results as those from variety 08127 were obtained. The proteomics profiling of seedlings in different ages of B. napus was analyzed by using the previous established protocol, we found 20 up-regulated and 51 down-regulated proteins with more than two times changes in 30 days old B. napus stems compared with in 15 days old B. napus stems and some of the different protein spots were successfully identified. The system established in this study is useful for B. napus proteomic research in the future.

Key words: Brassica napus, Protein, Two-dimensional polyacrylamide gel electrophoresis

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