Investigation of the effects of 2‐naphthylamine on the protein expression of Pseudomonas putida KT2440

Abstract

Two‐dimensional gel electrophoresis can be used in proteomics to study changes in protein expression. Currently, this tool is being employed to determine the impact of 2‐Naphthylamine, a known carcinogen, on the growth and protein expression of Pseudomonas putida KT2440. In order to grow strong cells and to increase gel resolution, modifications were made to growth media concentrations and protein isolation methods. Following this adapted procedure, a control was produced using Succinic Acid in Hutner's Growth media and new two‐dimensional control gels were analyzed. The optimal concentration at which to grow the bacteria in the presence of 2‐Naphthylamine was determined to be 0.25mM. The proteins from the 2‐Naphthylamine variable are similarly separated using two‐dimensional gel electrophoresis and the gels are analyzed using the PDQuest Software. A comparison is made between the 2‐napthylamine gels and the Succinic Acid control gels to look for changes in the bacteria's protein expression. Changes in protein expression in the 2‐Naphthylamine gels (when compared to the master control gel) can be new proteins, missing proteins, and under or over‐expression of proteins. Financial support by the Rochester Institute of Technology College of Science Dean's Office and the Department of Chemistry.