Abstract
The enhancement of protein-DNA interactions in the presence of glutamate salts has been observed in numerous studies and has often been called “the glutamate effect”. Klenow and Klentaq are the large fragment domains of the Type I DNA polymerases from E. coli and Thermus aquaticus, respectively. Our lab has previously characterized the enhancing effect of potassium glutamate (KGlu) on the the binding affinities of these polymerases to DNA versus that of potassium chloride (KCl). In this project, we investigate the effect of KGlu on the overall functionalities of Klenow and Klentaq.Replacing KCl with KGlu in the reaction buffer for the polymerase chain reaction (PCR) results in a significant increase in the quantity of replication product at low salt concentrations ( 1M) appears unrelated to nucleotide incorporation activity. Ongoing work includes examining the effects of KGlu versus KCl on the melting temperature of duplex DNA, and use of real-time PCR to more fully characterize the functional enhancement effect of glutamate in the overall DNA replication reaction. This work is supported by the NSF.
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