Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli

A Na+/H+ antiporter gene was isolated from alkaliphilic Bacillus sp. G1. The full-length sequence of the Na+/H+ antiporter gene was obtained using a genome walking method, and designated as g1-nhaC. An ORF preceded by a promoter-like sequence and a Shine—Dalgarno sequence, and followed by a termin...

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Main Authors: Chong, Wai Liew, Md. Illias, Rosli, Mahadi, Nor Muhammad, Najimudin, Nazalan
Format: Article
Language:English
Published: Blackwell Publishing Ltd. 2007
Subjects:
Online Access:http://eprints.utm.my/5903/
http://eprints.utm.my/5903/1/ChongWaiLiew2007_ExpressionOfTheNa%2BH%2BAntiporterGene.pdf
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author Chong, Wai Liew
Md. Illias, Rosli
Mahadi, Nor Muhammad
Najimudin, Nazalan
author_facet Chong, Wai Liew
Md. Illias, Rosli
Mahadi, Nor Muhammad
Najimudin, Nazalan
author_sort Chong, Wai Liew
building UTeM Institutional Repository
collection Online Access
description A Na+/H+ antiporter gene was isolated from alkaliphilic Bacillus sp. G1. The full-length sequence of the Na+/H+ antiporter gene was obtained using a genome walking method, and designated as g1-nhaC. An ORF preceded by a promoter-like sequence and a Shine—Dalgarno sequence, and followed by a terminator-like sequence was identified. The deduced amino acid sequence consists of 535 amino acids, and a calculated molecular mass of 57 776 Da. g1-nhaC was subsequently cloned into pET22b(+) and expressed in Escherichia coli BL21 (DE3). Recombinant E. coli harboring the g1-nhaC gene was able to grow in modified L medium at various concentrations of NaCl (0.2-2.0 M) at different pH values. The recombinant bacteria grew well in the medium with concentrations of NaCl as high as 1.75 M at pH 8.0-9.0. Minimal growth was observed at 2.0 M NaCI, pH 8.0-9.0. At pH 10, the recombinant bacteria grew well in a medium with a low concentration of NaCl (0.2 M). These results suggested that the g1-NhaC antiporter from Bacillus sp. G1 plays a role in Na+ extrusion at lower pH values and in pH homeostasis at pH 10 under Na+ -limiting conditions.
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spelling utm-59032010-06-01T15:36:23Z http://eprints.utm.my/5903/ Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli Chong, Wai Liew Md. Illias, Rosli Mahadi, Nor Muhammad Najimudin, Nazalan T Technology (General) A Na+/H+ antiporter gene was isolated from alkaliphilic Bacillus sp. G1. The full-length sequence of the Na+/H+ antiporter gene was obtained using a genome walking method, and designated as g1-nhaC. An ORF preceded by a promoter-like sequence and a Shine—Dalgarno sequence, and followed by a terminator-like sequence was identified. The deduced amino acid sequence consists of 535 amino acids, and a calculated molecular mass of 57 776 Da. g1-nhaC was subsequently cloned into pET22b(+) and expressed in Escherichia coli BL21 (DE3). Recombinant E. coli harboring the g1-nhaC gene was able to grow in modified L medium at various concentrations of NaCl (0.2-2.0 M) at different pH values. The recombinant bacteria grew well in the medium with concentrations of NaCl as high as 1.75 M at pH 8.0-9.0. Minimal growth was observed at 2.0 M NaCI, pH 8.0-9.0. At pH 10, the recombinant bacteria grew well in a medium with a low concentration of NaCl (0.2 M). These results suggested that the g1-NhaC antiporter from Bacillus sp. G1 plays a role in Na+ extrusion at lower pH values and in pH homeostasis at pH 10 under Na+ -limiting conditions. Blackwell Publishing Ltd. 2007 Article PeerReviewed application/pdf en http://eprints.utm.my/5903/1/ChongWaiLiew2007_ExpressionOfTheNa%2BH%2BAntiporterGene.pdf Chong, Wai Liew and Md. Illias, Rosli and Mahadi, Nor Muhammad and Najimudin, Nazalan (2007) Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli. FEMS Microbiology Letters, 276 . pp. 114-122.
spellingShingle T Technology (General)
Chong, Wai Liew
Md. Illias, Rosli
Mahadi, Nor Muhammad
Najimudin, Nazalan
Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli
title Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli
title_full Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli
title_fullStr Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli
title_full_unstemmed Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli
title_short Expression of the Na+/H+ antiporter gene (g1-nhaC) of alkaliphilic bacillus sp. G1 in escherichia coli
title_sort expression of the na+/h+ antiporter gene (g1-nhac) of alkaliphilic bacillus sp. g1 in escherichia coli
topic T Technology (General)
url http://eprints.utm.my/5903/
http://eprints.utm.my/5903/1/ChongWaiLiew2007_ExpressionOfTheNa%2BH%2BAntiporterGene.pdf