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Journal of Undergraduate Research

Keywords

twelve member plasmid library, promotor swapping, chromosomal gene expression in E. coli

College

Life Sciences

Department

Microbiology and Molecular Biology

Abstract

Metabolic engineering is becoming a very important area of research, allowing researchers to harness metabolic pathways to either eliminate or synthesize desired compounds. Understanding metabolic pathways by altering expression of proteins involved in the pathway helps to uncover thermodynamic bottlenecks which render certain pathways inefficient or infeasible. Altering expression of genes through the use of promoter swapping is a useful research technique used to understand important metabolic and physiologic pathways. Engineered promoters contained in plasmids can be inserted into chromosomal DNA through the utilization of promoter swapping to alter the expression of a given gene in order to better understand the pathway. We proposed creating a library of twelve plasmids containing promoter sequences capable of altering gene expression at varying, constitutive levels with an accompanying protocol for insertion of the engineered promoter sequence into the chromosome upstream of any gene of Escherichia coli. We have successfully synthesized two promoters which have altered β–galactosidase expression in cultured cells as desired.

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