Characterization of the Interaction of Escherichia coli dnaB and dnaC Proteins Using the Two-Hybrid System
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The purpose of this project was to develop a system to characterize the protein:protein interaction of E. coli replication proteins dnaB and dnaC. Experiments were done using the yeast Two-Hybrid system. In this system, the interaction of two proteins, each fused to a separate domain of a yeast transcriptional activator protein, can be measured by the reconstitution of functional transcription of a reporter gene in recombinant yeast. Results of these studies prove that these proteins interact specifically and with high affinity. This recombinant model system can be used to define the regions of dnaB and dnaC that interact by providing a facile assay in which the functional competence of mutated forms of each protein can be measured.