Production and characterization of MutS for use in error correction

Abstract

The availability of inexpensive synthetic DNA (oligonucleotides) has allowed for the synthesis of longer, gene-length constructs of DNA. However, a critical barrier to making this technology a low-cost and high-throughput process has been due to the rate at which errors pervade the final product. The current state of the error reduction technology includes three different categories: error filtration, error correction, and error prevention. My research is a joint project as well as an addendum to the work done by Research Scientist Dr. Peter Carr and current MIT Department of Biological Engineering Masters Student Jason Park (MIT '05) who have been working on research in gene synthesis error correction over the past several years. I have been working very closely with both Dr. Carr and Jason Park on this research for the past two years. We have a publication we're about to submit in regards to optimizations of gene synthesis and a significant portion of my thesis deals with work done for the upcoming publication. My work includes optimizing the synthesis of large gene constructs, the synthesis of new hyper-thermophilic MutS proteins, characterizing these proteins using instruments such as the circular dichroism spectrophotometer and the Evotec MF20, as well as perfecting old error correction protocols while designing several new ones.