Proteomic studies of pseudomonas putida KT2440 in carcinogenicity screening via 2-dimensional gel electrophoresis

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Title: Proteomic studies of pseudomonas putida KT2440 in carcinogenicity screening via 2-dimensional gel electrophoresis
Author: Yao, Mingyi
Abstract: This project is part of a pilot study designed to establish a two dimensional gel electrophoresis-based protocol, which can be utilized in screening mutagencity/carcinogencity of uncharacterized substances. Currently, the Ames test is the most commonly used method, which is solely based on mutation at the genome level. In contrast to the genome, which is static, protein expression in a cell under a given set of conditions (the proteome) is dynamic. A screening method based on the proteome rather than the genome may prove more reliable and quantitative in predicting mutagenicity and carcinogenicity. In this project, Pseudomonas putida KT2440 was employed as the biological model because its genome has been completely sequenced and the strain is already known to be able to grow on several aromatic carbon sources. Two carbon sources were selected, succinic acid and phenylethylamine. The strain grew in the non-aromatic and aromatic carbon source respectively. Proteins were extracted at a fixed growth stage (mid-log phase) so that protein expression is consistent. Protein expression was then analyzed by two dimensional gel electrophoresis, with the hope of identifying proteins that are expressed in response to phenylethylamine ? the proteomic signature for phenylethylamine. In total, six signature protein spots were found. In addition to identifying these signature protein spots, a major goal of this project (to establish standard protocols for protein expression and analysis) was accomplished. Based on the results obtained in this project, several directions for further improvement in later stages of the procedure are also discussed.
Record URI: http://hdl.handle.net/1850/1118
Date: 2005-11-02

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