Establishment of Two-dimensional Electrophoresis System of Tomato Cotyledons

ACTA HORTICULTURAE SINICA ›› 2010, Vol. 37 ›› Issue (4): 661-668.

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Establishment of Two-dimensional Electrophoresis System of Tomato Cotyledons

LI Fei1，XU Qiu-fang1，ZANG Xian-peng1，LAI Yi-yu1，CHENG Wei-shun1，XU You-ping2,*，and CAI Xin-zhong1,*

（1Institute of Biotechnology，College of Agriculture and Biotechnology，Zhejiang University，Hangzhou 310029，China；2Centre of Analysis and Measurement，Zhejiang University，Hangzhou 310029，China）

Received:2009-09-22 Revised:2010-03-23 Online:2010-04-25 Published:2010-04-25
Contact: XU You-ping,CAI Xin-zhong

Abstract

Abstract: A two-dimensional electrophoresis（2-DE）system for proteomic analysis of tomato cotyledons was established by optimizing the parameters including the pH gradient of IPG strip，sample application，sample loading and isoelectric focusing conditions. The optimized system include the following steps：Extracting the total protein from tomato cotyledons by TCA/acetone precipitation，separating the proteins with 24 cm pH3-7NL IPG strips，loading protein samples of 300 μg by rehydration method followed by isoelectric focusing programⅠorⅡ，and staining the gels by silver staining after SDS-PAGE electrophoresis. When preparative gel was applied，in order to obtain abundant protein spots，1.5 mg sample was loaded followed by the isoelectric focusing program Ⅲ，and staining with colloidal Coomassie Brilliant Blue. Reproducible profiles with high resolution were obtained by this optimized 2-DE system of tomato cotyledon.

Key words: tomato, two-dimensional electrophoresis（2-DE）, condition optimization, cotyledon, protein

CLC Number:

S 641.2

LI Fei;XU Qiu-fang;ZANG Xian-peng;LAI Yi-yu;CHENG Wei-shun;XU You-ping;and CAI Xin-zhong;. Establishment of Two-dimensional Electrophoresis System of Tomato Cotyledons[J]. ACTA HORTICULTURAE SINICA, 2010, 37(4): 661-668.

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Establishment of Two-dimensional Electrophoresis System of Tomato Cotyledons

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