An Efficient Method for Extraction of Secreted Proteins of a Filamentous Fungus, Cryphonectria parasitica
Jinzi Wang, Fangzhen Wang, Jinjie Shang and Baoshan Chen*
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Science and Technology, Guangxi University, Nanning 530004, China
- *Corresponding Author:
- Dr. Baoshan Chen
College of Life Science and Technology
Guangxi University, Nanning, 530004, PR China
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Received Date: May 09, 2011; Accepted Date: June 17, 2011; Published Date: June 20, 2011
Citation: Wang J, Wang F, Shang J, Chen B (2011) An Efficient Method for Extraction of Secreted Proteins of a Filamentous Fungus, Cryphonectria parasitica. J Proteomics Bioinform 4: 125-128. doi:10.4172/jpb.1000179
Copyright: © 2011 Wang J, 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.
Three different methods were evaluated for extraction of secreted proteins from a filamentous fungus, Cryphonectria parasitica, cultured in liquid medium. Protein samples prepared with each method were subjected to SDS-PAGE and two-dimensional gel electrophoresis (2-DE). It was found that ultrafiltration was not suitable for extraction of the secreted proteins. Protein samples prepared by chloroform treatment followed by ammonium sulfate precipitation were acceptable for 2-DE separation. The modified sevage method was the best to extract secreted proteins with highest yield and best quality. About 300 protein spots could be identified on a 2-DE gel using protein samples prepared with this method from 3 day-old culture. Successful isolation of high quality secreted proteins is the pre-request for the analysis of fungal secreted proteome. The new method may be applied to other filamentous fungi, particularly the pathogenic ones.