The growing problem of antibiotic resistance has prompted the need for new and
novel antimicrobial therapies. The bacterial cell division pathway holds great promise for
the development of novel broad-spectrum antibiotics as the majority of the proteins are
essential for viability. The wealth of information regarding bacterial cell division has
come from studies of the model organisms Escherichia coli and Bacillus subtilis.
Although much has been elucidated regarding this pathway, the functions of many
individual proteins remain unsolved. An important model organism for the investigation of cell division is Streptomyces coelicolor. The mycelial Streptomyces are sporulating, Gram-positive
bacteria that grow in long branching networks of filamentous cells much like filamentous
fungi. The normally essential process of cell division is dispensable for growth and
viability of S. coelicolor. More interestingly, there are two different modes of cell
division in this organism, one for vegetative growth and one is utilized for synchronous
septation during sporulation. It is still unclear how developmental regulators control this
switch, but advancements in fluorescence microscopy have shed some insight into the cell
division process by allowing direct visualization of many cellular components and their
dynamics. To better understand bacterial cell division and its regulation in S. coelicolor,
three additional fluorescent proteins (FPs), including m.RFP, CyPet and YPet, have been
established in this work. An m.RFP shuttle vector was constructed and the utility of
m.RFP was tested by translationally fusing it to a tip-localizing protein, DiviVA. This
work demonstrated that m.RFP is functional and an efficient marker for localized proteins.
Also, established in this work is a two-colour fluorescence reporter system, which
includes the fluorescent proteins CyPet and YPet that can be used to study co-localization
and protein-protein interactions within cells. Future plans are to use co-localization of FP
fusions and fluorescence resonance energy transfer (FRET) between CyPet and YPet to
investigate the assembly of protein complexes within the cells, such as those involved in
cell division. These studies will reveal critical information that is needed for the
development of drugs that have novel mechanisms of action. / Thesis / Master of Science (MSc)
| Identifer | oai:union.ndltd.org:mcmaster.ca/oai:macsphere.mcmaster.ca:11375/21608 |
| Date | 09 1900 |
| Creators | Nguyen, Khoa |
| Contributors | Nodwell, Justin, Biochemistry and Biomedical Sciences |
| Source Sets | McMaster University |
| Language | English |
| Detected Language | English |
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