Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp

The development of sensitive methods for observing individual bacterial cells in a population in experiment and natural environments is very crucial for rapid development of anti-mycobacterial drugs. This is because more pathogenic Mycobacteria are slow growing organisms, and therefore the screen...

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Main Author:	Najamudin, Khairal Ezani
Format:	Thesis
Language:	English
Published:	2003
Subjects:	RC Internal medicine
Online Access:	http://eprints.usm.my/44952/1/PTA...Khairul%20Ezani%20Najamudin...2003...-24%20pages.pdf
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id	my-usm-ep.44952
record_format	uketd_dc
spelling	my-usm-ep.449522020-10-22T03:03:26Z Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp 2003 Najamudin, Khairal Ezani RC Internal medicine The development of sensitive methods for observing individual bacterial cells in a population in experiment and natural environments is very crucial for rapid development of anti-mycobacterial drugs. This is because more pathogenic Mycobacteria are slow growing organisms, and therefore the screening compounds for anti-mycobacterial activity is slow and inefficient. Previously, studies have been done using fluorescent bacteria that expressing 13- galactosidase (21 ), and luciferase (3) as a high screening format for antimicrobial activity. However, one drawback of these systems is that substrates such as luciferin have to be added at the required time points to induce fluorescence. Recently Green Fluorescent Protein (GFP) has become a valuable and favourite tool as a marker of growth, which could be used for screening of antimicrobial activity. This is because the marker can be visualised without interruption or termination of an experiment as is required with the detection of other commonly used markers such as B-galactosidase and luciferase. In this study a synthetic gene of GFP was constructed with mycobacteria codon bias using assembly PCR. A strong mycobacterial promoter from 65 kD mycobacterial heat shock protein was added to derive the expression. The constructed gene was cloned into vector and transformed into Escherichia coli. The ultimate aim is to use the recombinant plasmid carrying the synthetic gene with mycobacterial codon bias to create a recombinant fluorescing BCG, whichcan be used as a tool for screening compound libraries for anti-mycobacterial activity. 2003 Thesis http://eprints.usm.my/44952/ http://eprints.usm.my/44952/1/PTA...Khairul%20Ezani%20Najamudin...2003...-24%20pages.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Kesihatan
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	RC Internal medicine
spellingShingle	RC Internal medicine Najamudin, Khairal Ezani Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp
description	The development of sensitive methods for observing individual bacterial cells in a population in experiment and natural environments is very crucial for rapid development of anti-mycobacterial drugs. This is because more pathogenic Mycobacteria are slow growing organisms, and therefore the screening compounds for anti-mycobacterial activity is slow and inefficient. Previously, studies have been done using fluorescent bacteria that expressing 13- galactosidase (21 ), and luciferase (3) as a high screening format for antimicrobial activity. However, one drawback of these systems is that substrates such as luciferin have to be added at the required time points to induce fluorescence. Recently Green Fluorescent Protein (GFP) has become a valuable and favourite tool as a marker of growth, which could be used for screening of antimicrobial activity. This is because the marker can be visualised without interruption or termination of an experiment as is required with the detection of other commonly used markers such as B-galactosidase and luciferase. In this study a synthetic gene of GFP was constructed with mycobacteria codon bias using assembly PCR. A strong mycobacterial promoter from 65 kD mycobacterial heat shock protein was added to derive the expression. The constructed gene was cloned into vector and transformed into Escherichia coli. The ultimate aim is to use the recombinant plasmid carrying the synthetic gene with mycobacterial codon bias to create a recombinant fluorescing BCG, whichcan be used as a tool for screening compound libraries for anti-mycobacterial activity.
format	Thesis
qualification_level	Master's degree
author	Najamudin, Khairal Ezani
author_facet	Najamudin, Khairal Ezani
author_sort	Najamudin, Khairal Ezani
title	Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp
title_short	Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp
title_full	Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp
title_fullStr	Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp
title_full_unstemmed	Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp
title_sort	construction of the aequorea victoria green fluorescent protein (gfp) for expression in mycobacterium sp
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Kesihatan
publishDate	2003
url	http://eprints.usm.my/44952/1/PTA...Khairul%20Ezani%20Najamudin...2003...-24%20pages.pdf
_version_	1747821429458468864

Construction of the aequorea Victoria green fluorescent protein (gfp) for expression in mycobacterium sp

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