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dc.contributor.advisor Kreuzer, Kenneth en_US
dc.contributor.author Maduike, Nkabuije Zikaodi en_US
dc.date.accessioned 2012-09-04T13:17:33Z
dc.date.issued 2012 en_US
dc.identifier.uri http://hdl.handle.net/10161/5863
dc.description Dissertation en_US
dc.description.abstract <p>Constitutive stable DNA replication (cSDR) is an alternative mode of replication initiation in Escherichia coli. cSDR is active in rnhA and recG mutants, which lack proteins that remove DNA-RNA hybrids called R-loops. The mechanism for cSDR initiation, therefore, is thought to involve these R-loop structures, which are proposed to form at specific locations known as oriK sites on the chromosome. Thus far, oriK sites have only been mapped to broad, 100-200 kb regions on the chromosome, so the specific elements involved in initiation are still unknown. My research focused on localizing the oriK sites on the chromosome, specifically those in the terminus region, where two of the major oriK sites had previously been mapped. We used two-dimensional gel electrophoresis (Friedman & Brewer, 1995) to analyze the replication forks that are blocked at the innermost Ter sites at the terminus, and found that elevated levels of replication forks are blocked at the Ter sites in rnhA mutants. We also used microarray and deep sequencing analysis to determine that there is a major location of oriK activity in the chromosome, located in the region between TerA and TerC. Furthermore, we also studied the use of the activation-induced deaminase (AID) enzyme as a tool for identifying regions of R-loop formation in the chromosome, and learned about its properties in the process.</p> en_US
dc.subject Biochemistry en_US
dc.subject cSDR en_US
dc.subject oriK en_US
dc.subject Replication en_US
dc.subject R-loops en_US
dc.title Native Origins for Constitutive Stable DNA Replication in Escherichia Coli en_US
dc.type Dissertation en_US
dc.department Biochemistry en_US
duke.embargo.months 24 en_US
duke.embargo.release 2014-08-25

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