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KMID : 0880220080460040415
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Journal of Microbiology
2008 Volume.46 No. 4 p.415 ~ p.421
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Cloning and Characterization of a Na+/H+ Antiporter Gene of the Moderately Halophilic Bacterium Halobacillus aidingensis AD-6T
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Zou Ya-Jie
Yang Li-Fu
Wang Lei
Yang Su-Sheng
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Abstract
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A gene encoding a Na+/H+ antiporter was obtained from the genome of Halobacillus aidingensis AD-6T, which was sequenced and designated as nhaH. The deduced amino acid sequence of the gene was 91% identical to the NhaH of H. dabanensis, and shared 54% identity with the NhaG of Bacillus subtilis. The cloned gene enable the Escherichia coli KNabc cell, which lack all of the major Na+/H+ antiporters, to grow in medium containing 0.2 M NaCl or 10 mM LiCl. The nhaH gene was predicted to encode a 43.5 kDa protein (403 amino acid residues) with 11 putative transmembrane regions. Everted membrane vesicles prepared from E. coli KNabc cells carrying NhaH exhibited Na+/H+ as well as Li+/H+ antiporter activity, which was pH-dependent with the highest activity at pH 8.0, and no K+ /H+ antiporter activity was detected. The deletion of hydrophilic C-terminal amino acid residues showed that the short C-terminal tail was vital for Na+/H+ antiporter activity.
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KEYWORD
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Halobacillus aidingensis, Na+/H+ antiporter, nhaH, Inverse PCR
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