Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model

Internalization and intracellular proliferation of pathogenic bacteria Leptospira interrogans (L. interrogans) in their host is induced by pathogenic proteins known as virulence factors (VFs) which target molecular mechanisms, cellular processes and eukaryotic structures that are conserved among euk...

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Main Author:	Lai, Weng Yu
Format:	Thesis
Language:	English
Published:	2019
Subjects:	QH1-278.5 Natural history (General)
Online Access:	http://eprints.usm.my/48641/1/190131_LWY%20edited%20thesis-EOedit%20cut.pdf
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id	my-usm-ep.48641
record_format	uketd_dc
spelling	my-usm-ep.486412021-03-17T02:40:15Z Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model 2019-02 Lai, Weng Yu QH1-278.5 Natural history (General) Internalization and intracellular proliferation of pathogenic bacteria Leptospira interrogans (L. interrogans) in their host is induced by pathogenic proteins known as virulence factors (VFs) which target molecular mechanisms, cellular processes and eukaryotic structures that are conserved among eukaryotes. Yeast, Saccharomyces cerevisiae (S. cerevisiae), is a popular eukaryotic model that is used to identify bacterial VFs. The goal of this study was to identify L. interrogans putative virulence factors by expressing them in yeast and examining the putative VFs impact on yeast growth. A total of 288 L. interrogans putative virulence genes were selected through a bioinformatic approach and literature review. Of the 288 selected putative virulence genes, 226 putative virulence genes were successfully cloned in the yeast expression plasmid and expressed in yeast using a 96-well plate format. The cell viability of yeast expressing putative virulence factors was verified using a liquid medium method and yeast serial dilution spotting assay. Eleven L. interrogans putative VFs were found inhibiting yeast growth and they were investigated for their localization in yeast and VF-yeast protein interaction. A green fluorescent protein (GFP) was fused to the C-terminal of the VFs to investigate the localization of putative VFs in yeast with the use of a fluorescent microscope. In conclusion, the yeast model is a simple and effective model to identify bacterial VFs that target conserved molecular mechanisms and cellular processes among yeast and human cells. This study has provided some preliminary information for the future study on L. interrogans invasion and evasion mechanisms. 2019-02 Thesis http://eprints.usm.my/48641/ http://eprints.usm.my/48641/1/190131_LWY%20edited%20thesis-EOedit%20cut.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Kajihayat
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	QH1-278.5 Natural history (General)
spellingShingle	QH1-278.5 Natural history (General) Lai, Weng Yu Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model
description	Internalization and intracellular proliferation of pathogenic bacteria Leptospira interrogans (L. interrogans) in their host is induced by pathogenic proteins known as virulence factors (VFs) which target molecular mechanisms, cellular processes and eukaryotic structures that are conserved among eukaryotes. Yeast, Saccharomyces cerevisiae (S. cerevisiae), is a popular eukaryotic model that is used to identify bacterial VFs. The goal of this study was to identify L. interrogans putative virulence factors by expressing them in yeast and examining the putative VFs impact on yeast growth. A total of 288 L. interrogans putative virulence genes were selected through a bioinformatic approach and literature review. Of the 288 selected putative virulence genes, 226 putative virulence genes were successfully cloned in the yeast expression plasmid and expressed in yeast using a 96-well plate format. The cell viability of yeast expressing putative virulence factors was verified using a liquid medium method and yeast serial dilution spotting assay. Eleven L. interrogans putative VFs were found inhibiting yeast growth and they were investigated for their localization in yeast and VF-yeast protein interaction. A green fluorescent protein (GFP) was fused to the C-terminal of the VFs to investigate the localization of putative VFs in yeast with the use of a fluorescent microscope. In conclusion, the yeast model is a simple and effective model to identify bacterial VFs that target conserved molecular mechanisms and cellular processes among yeast and human cells. This study has provided some preliminary information for the future study on L. interrogans invasion and evasion mechanisms.
format	Thesis
qualification_level	Master's degree
author	Lai, Weng Yu
author_facet	Lai, Weng Yu
author_sort	Lai, Weng Yu
title	Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model
title_short	Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model
title_full	Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model
title_fullStr	Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model
title_full_unstemmed	Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model
title_sort	identification of putative virulence factors of leptospira interrogans using a yeast model
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Kajihayat
publishDate	2019
url	http://eprints.usm.my/48641/1/190131_LWY%20edited%20thesis-EOedit%20cut.pdf
_version_	1747821955263758336

Identification Of Putative Virulence Factors Of Leptospira Interrogans Using A Yeast Model

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