M.S. selective enrichment of rare mutations as a new biotechnology to study DNA mismatch repair processes in bacteria

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)

Parth H. Desai (Creator)

Institution

The University of North Carolina at Greensboro (UNCG )

Web Site: http://library.uncg.edu/

Advisor

Eric Josephs

Abstract: DNA mismatch repair (MMR) is the key process which ensures the incorporation of correct nucleotides during DNA replication by recognizing and removing of incorrectly paired nucleotides from DNA. DNA replication can introduce a mismatched nucleotide at a rate of 10-5 to 10-6 nts per replication cycle. If this mismatch is not corrected, then it becomes a permanent mutation after the next round of replication. Understanding the MMR mechanism can yield important insights into many aspects of human health, like the emergence of cancer and drug resistance in bacteria. To overcome experimental challenges with studying this process in living cells, we have developed a new method to enrich the rare genomic mutations by genotypic selection in a way that allows us to study the mismatch repair process in Escherichia coli, a model organism for MMR. We have shown the maximum 705,000-fold enrichment of DNA with a mutation even after a 10-6 times dilution by DNA with the wild-type sequence in vitro. After further optimization, we could then use this technique to directly measure MMR activity occurring in living E. coli (in vivo). We expect this technique will open up new opportunities and research directions to study MMR-like processes in E. coli as well as different organisms, including Actinobacteria.

M.S. selective enrichment of rare mutations as a new biotechnology to study DNA mismatch repair processes in bacteria
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Created on 5/1/2020
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Additional Information

Publication

Thesis

Language: English

Date: 2020

Keywords

Bacteria, Biotechnologies, Mismatch Repair, Rare Mutations, Study

Subjects

DNA repair

Microbial mutation

Biotechnology

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