Use of a synthetic riboswitch to determine the effects of Erc on the regulatory element CovR and associated virulence genes in Streptococcus pyogenes Público
Baecher, Kirsten Mary (2011)
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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