alexa Optimization of Two-Dimensional Gel Electrophoresis Analysis of Chinese Cabbage (Brassica rapa L. Ssp. Pekinensis) Buds | OMICS International | Abstract
ISSN: 2376-0354

Journal of Horticulture
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Research Article

Optimization of Two-Dimensional Gel Electrophoresis Analysis of Chinese Cabbage (Brassica rapa L. Ssp. Pekinensis) Buds

Xue Zhou, Ruiqin Ji, Zhiyong Liu and Hui Feng*
Department of Horticulture, Shenyang Agricultural University, Shenyang 110866, China
Corresponding Author : Hui Feng
Department of Horticulture
Shenyang Agricultural University
Shenyang 110866, China
Tel: 86-13591688831
E-mail: [email protected]
Received: April 01, 2015 Accepted: June 14, 2015 Published: June 17, 2015
Citation: Zhou X, Ji R, Liu Z, Feng H (2015) Optimization of Two-Dimensional Gel Electrophoresis Analysis of Chinese Cabbage (Brassica rapa L. Ssp. Pekinensis) Buds. J Horticulture 2:146. doi:10.4172/2376-0354.1000146
Copyright: © 2015 Zhou X, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Abstract

In order to establish a suitable and effective protocol for two-dimensional gel electrophoresis (2-DE) analysis of the buds of Brassica rapa L. ssp. Pekinensis, whole proteins from fertile buds of the A/B line AB01 were extracted using three different methods (Tris-acetone-phenol method, Trizol precipitation method, TCA-acetone precipitation method). Furthermore, four aspects of the 2-DE system were optimized, including the IEF protocol, SDS-PAGE gel concentration, pH range of IPG strips and protein sample volume. The results show that the TCA-acetone precipitation method is the most effective method for whole protein extraction, and an optimized 2-DE system using the 20,000 Vh IEF protocol, pH 4-7 IPG strips, 12% SDS-PAGE gel concentration and protein sample volumes of 250 μg is suitable for 2-DE analysis of fertile bud protein in Chinese cabbage. These results can be used to facilitate proteomic studies of genic male sterility in Brassica rapa L. ssp. Pekinensis.

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