Use of a synthetic riboswitch to determine the effects of Erc on the regulatory element CovR and associated virulence genes in Streptococcus pyogenes Open Access

Baecher, Kirsten Mary (2011)

Permanent URL: https://etd.library.emory.edu/concern/etds/8336h2360?locale=en
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Abstract

Abstract
Use of a synthetic riboswitch to determine the effects of Erc on the regulatory element CovR and
associated virulence genes in Streptococcus pyogenes
By Kirsten Baecher
The human pathogen Streptococcus pyogenes (Group A Streptococcus, GAS) is responsible for
various diseases, which range in severity from minor self-clearing infections like those of the
throat and skin to severe, life-threatening diseases such as necrotizing fasciitis. The severity of an
infection is mediated by the expression of virulence factors encoded in the S. pyogenes
chromosome. One mechanism by which expression of these factors (as well as other genes) is
regulated is the 2-component regulatory system, CovR/S. CovR is a response regulator protein
that, when bound to DNA, represses transcription. It has been demonstrated that in addition to
being under control of its own promoter, CovR transcription is also promoted by the upstream
Erc promoter. The role of the Erc protein in this pathway, however, remains unclear. In this
investigation, I worked towards construction of a plasmid that could be used to control Erc
translation via a synthetic riboswitch, which would ultimately be used in the host chromosome
for assays of expression of elements under CovR control, at different levels of Erc expression. I
discuss challenges that were presented during the construction of the plasmid and their molecular
basis, as well as potential strategies for overcoming them in future work.

Table of Contents

Table of Contents


Index of Figures……………………………………………………………....i
I.Purpose………………………………………………………………………...1
II.Introduction………………………………………………………………….1
III.Methods……………………………………………………………………...6
IV.Results………………………………………………………………………...6
V.Discussion…………………………………………………………………...14
VI.References …………………………………………………………………17

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