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Journal of Tea Science ›› 2013, Vol. 33 ›› Issue (5): 420-428.doi: 10.13305/j.cnki.jts.2013.05.012

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Establishment of Two-dimensional Electrophoresis System for Pistil Proteome Analysis in Tea Plant

REN Yan1,2, CHEN Xuan1, FANG Wan-ping1, LI Yao1, LI Xing-hui1,2,*   

  1. 1. College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China;
    2. Suzhou Engineering Research Center for Modern Ecological Tea Industry, Suzhou 215128, China
  • Received:2013-04-22 Revised:2013-07-27 Online:2013-10-30 Published:2019-09-04

Abstract: In order to establish a proper two-dimensional gel electrophoresis (2-DE) protocol for proteomic study of tea plant pistils, several protein extraction methods, density of SDS-PAGE gel, IEF procedures and pH gradient of IPG strip were compared. The results showed the optimized system includes: total proteins of sample extracted using TCA/acetone method followed with clean up, separating the proteins with 17βcm pH 5~8 IPG strips, 13% SDS-PAGE, and CBB R-250 stain method. About 1β200 spots can be detected, and these proteins mainly distributed in the pH 5~8, MW 14~117βkD range. With good separation of the basic protein, this will be helpful to proteomic research in tea plant.

Key words: Camellia sinensis (L.) O. Kuntze, pistil, protein sample preparation, two-dimensional electrophoresis

CLC Number: