DNA polymerases (DNAp) catalyze the correct replication and repair of DNA, an essential step in the life cycle of all organisms. The high fidelity of DNAp requires an intricate mechanism to ensure correct nucleotide (nt) incorporation. Single-molecule studies can identify the dynamics of DNAp structural conformations, which are otherwise lost through averaging in ensemble populations. Therefore, the Weiss and Collins labs (UCI) collaboratively developed a single-walled carbon nanotube field-effect transistor (SWCNT-FET) to translate enzyme motions into electronic signals. These SWCNT-FETs may illuminate hidden conformational transitions of DNAp activity and reveal new mechanisms and dynamics during DNA synthesis. The goal of this project is to understand the enzyme dynamics of the thermostable DNAp from <i>Thermus aquaticus </i>(Taq) at single molecule level. This work may resolve the subtle and transient intermediate states of Taq that modulate catalytic speed and molecular recognition during DNA synthesis. Our efforts are focused on evaluating the performance of SWCNT-FETs to increase potential DNA sequencing capabilities.

Dissecting DNA polymerase at the single-molecule level

2019 Fall National Meeting

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