Title

Characterizing a homolog of FEN-1 in Methanosarcina acetivorans: an investigation into the evolution of the DNA repair pathways of archaea

Document Type

Thesis

Publication Date

2012

Abstract

Saccharomyces cerevisiae has been used as a model organism for investigating the DNA repair pathways in eukaryotes for many years, and study of that organism has provided scientists with certain understandings of how different eukaryotic organisms repair lesions on their DNA. However, to date little investigation has been done on the characterization of the DNA repair mechanisms of the most recently discovered domain of life, the archaea. This investigation used S. cerevisiae as a model organism for the investigation of a sequence homolog of Flap Endonuclease-1 in the archaea Methanosarcina acetivorans using recombinant DNA techniques. The gene for the archaeal FEN-1 was inserted into a strain of S. cerevisiae that had its FEN-1 equivalent knocked out and tests were run to determine the level of complementation the archaeal protein provided in the knockout yeast. It was determined that the FEN-1 from M. acetivorans was capable of partial complementation of wild-type function in the knockout yeast. With this observed complementation, suggestions can be made as to the development of eukaryotic-like DNA repair mechanisms in archaea. Furthermore, through this study, the groundwork has been laid for additional research into the DNA metabolism of the third domain of life.

Comments

Under the advisement of Michael Reagan.

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