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High Throughput Studies of Gene Expression Using Green Fluorescent Protein-Oxidative Stress Promoter Probe Constructs The Potential for Living Chips

Molecular and Cellular Biology Program, University of Maryland Baltimore County, Baltimore, MD

Canghai Lu

Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County, Baltimore, MD

William E. Bentley

Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, and Department of Chemical Engineering, University of Maryland, College Park, College Park, MD

Govind Rao

Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County, Baltimore, MD, Medical Biotechnology Center of the Maryland Biotechnology Institute, University of Maryland, Baltimore, MD

Green fluorescent protein fusions were constructed with several oxidative stress promoters from Escherichia coli. These promoters were chosen for their induction by reactive oxygen species (ROS) such as superoxide, hydrogen peroxide, and hydroxyl radicals. When exposed to various free radical insults, the cells fluoresced with great specificity based on the corresponding ROS. In this work, we propose a way in which these constructs could be used to study the mode of action of a variety of antitumor drugs. This approach offers the possibility of complementing gene chip technology by the creation of living chips for high throughput screening as well as studying differential gene expression.

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