Establishment and Optimization of Two-dimensional Electrophoresis System of Watermelon Root

doi:10.7668/hbnxb.2015.S1.009

Abstract

Abstract: An efficient and suitable two-dimensional electrophoresis(2-DE) system for proteomic analysis of watermelon roots was established by optimizing the isoelectric focusing conditions,sample loading,the pH gradient of IPG strips and staining methods.Results showed that the optimized system for watermelon roots were as following steps:total protein of watermelon roots was extracted by TCA/acetone precipitation,lysis Buffer contained 7 mol/L Urea,2 mol/L Thouirea,4% CHAPS(m/V),1% Cocktail(V/V),65 mmol/L DTT and 2% pH 4-7 IPG Buffer,13 cm pH 4-7 IPG strips was used to separate the proteins,loading protein samples was 800 μg,IEF condition was 48 000 Vh,12.5% gel concentration and followed by staining with colloidal Coomassie Brilliant Blue R-250.

Key words: Watermelon roots, Proteomics, Two-dimensional electrophoresis

CLC Number:

Q78
S651.03

ZHANG Man, YANG Xing-ping, XU Jin-hua, LIU Guang, YAO Xie-feng, LI Ping-fang. Establishment and Optimization of Two-dimensional Electrophoresis System of Watermelon Root[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(S1): 51-54. doi: 10.7668/hbnxb.2015.S1.009.

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http://www.hbnxb.net/EN/Y2015/V30/IS1/51

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