Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea

Halogenated organic compound are widely used in agriculture and industry over the past 100 years. The use of these compounds in the environment led to human health problems and environmental pollution because of their persistence and toxicity. Dalapon or 2,2-dichloropropionic acid is widely used as...

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Main Author:	Rizan, Nastaran
Format:	Thesis
Language:	English
Published:	2013
Subjects:	QH301 Biology
Online Access:	http://eprints.utm.my/id/eprint/78289/1/NastaranRizanMFBME20131.pdf
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spelling	my-utm-ep.782892018-08-03T08:47:08Z Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea 2013-08 Rizan, Nastaran QH301 Biology Halogenated organic compound are widely used in agriculture and industry over the past 100 years. The use of these compounds in the environment led to human health problems and environmental pollution because of their persistence and toxicity. Dalapon or 2,2-dichloropropionic acid is widely used as herbicides and plant growth regulator which cause environmental pollution. In this research, bacterium NR1 was isolated from soil sample taken from Bandar-e Anzali seashore in north of Iran. The result has shown that this bacterium grow in minimal media containing 20mM 2, 2-DCP with doubling time of 0.89 hours, which indicated its ability to degrade 2, 2-DCP. Based on microscopic observation and Gram staining, strain NR1 was identified as rod Gram positive bacterium. Biochemical tests for the bacterium NR1 were positive for oxidase, catalase, gelatin liquefaction, nitrate reduction, TSI, oxidation- fermentation glucose, starch and casein test, while the tests result were negative for lactose utilization, citrate, indole, and urease test. Genomic DNA from bacterium NR1 was extracted and 16S rRNA PCR amplification was carried out using universal primers, Fd1 (5‟ - AGA GTT TGA TCC TGGCTC AG - 3‟) and rP1 (5‟- ACG GTC ATA CCT TGT TAC GAC TT - 3‟) before sending for sequencing. NR1 strain 16S rRNA sequences were applied for Basic Local Alignment Search Tool nucleotide (BLASTn) and further analyzed using phylogenetic tree of Neighbour-Joining method (MEGA 5). Phylogenetic analysis indicated that NR1 strain clearly shared 97% homology to the genus of Bacillus cereus and therefore designated as Bacillus cereus sp. NR1. The PCR analysis of dehalogenase using dhlb_F_314/dhlB_R_637 primers showed a band with approximate size of >200 bp, suggesting this bacterium carries dehalogenase from class I. 2013-08 Thesis http://eprints.utm.my/id/eprint/78289/ http://eprints.utm.my/id/eprint/78289/1/NastaranRizanMFBME20131.pdf application/pdf en public http://dms.library.utm.my:8080/vital/access/manager/Repository/vital:79542 masters Universiti Teknologi Malaysia, Faculty of Biosciences and Medical Engineering Faculty of Biosciences and Medical Engineering
institution	Universiti Teknologi Malaysia
collection	UTM Institutional Repository
language	English
topic	QH301 Biology
spellingShingle	QH301 Biology Rizan, Nastaran Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea
description	Halogenated organic compound are widely used in agriculture and industry over the past 100 years. The use of these compounds in the environment led to human health problems and environmental pollution because of their persistence and toxicity. Dalapon or 2,2-dichloropropionic acid is widely used as herbicides and plant growth regulator which cause environmental pollution. In this research, bacterium NR1 was isolated from soil sample taken from Bandar-e Anzali seashore in north of Iran. The result has shown that this bacterium grow in minimal media containing 20mM 2, 2-DCP with doubling time of 0.89 hours, which indicated its ability to degrade 2, 2-DCP. Based on microscopic observation and Gram staining, strain NR1 was identified as rod Gram positive bacterium. Biochemical tests for the bacterium NR1 were positive for oxidase, catalase, gelatin liquefaction, nitrate reduction, TSI, oxidation- fermentation glucose, starch and casein test, while the tests result were negative for lactose utilization, citrate, indole, and urease test. Genomic DNA from bacterium NR1 was extracted and 16S rRNA PCR amplification was carried out using universal primers, Fd1 (5‟ - AGA GTT TGA TCC TGGCTC AG - 3‟) and rP1 (5‟- ACG GTC ATA CCT TGT TAC GAC TT - 3‟) before sending for sequencing. NR1 strain 16S rRNA sequences were applied for Basic Local Alignment Search Tool nucleotide (BLASTn) and further analyzed using phylogenetic tree of Neighbour-Joining method (MEGA 5). Phylogenetic analysis indicated that NR1 strain clearly shared 97% homology to the genus of Bacillus cereus and therefore designated as Bacillus cereus sp. NR1. The PCR analysis of dehalogenase using dhlb_F_314/dhlB_R_637 primers showed a band with approximate size of >200 bp, suggesting this bacterium carries dehalogenase from class I.
format	Thesis
qualification_level	Master's degree
author	Rizan, Nastaran
author_facet	Rizan, Nastaran
author_sort	Rizan, Nastaran
title	Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea
title_short	Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea
title_full	Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea
title_fullStr	Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea
title_full_unstemmed	Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea
title_sort	screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea
granting_institution	Universiti Teknologi Malaysia, Faculty of Biosciences and Medical Engineering
granting_department	Faculty of Biosciences and Medical Engineering
publishDate	2013
url	http://eprints.utm.my/id/eprint/78289/1/NastaranRizanMFBME20131.pdf
_version_	1747817953132281856

Screen a dehalogenase gene using isolated bacterium from sea-shore soil of caspian sea

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