Cloning and Characterization of a Na+/H+ Antiporter Gene of the Moderately Halophilic Bacterium Halobacillus aidingensis AD-6T

Ya Jie Zou; Li Fu Yang; Lei Wang; Su Sheng Yang

doi:10.1007/s12275-008-0009-2

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Cloning and Characterization of a Na+/H+ Antiporter Gene of the Moderately Halophilic Bacterium Halobacillus aidingensis AD-6T: Ya Jie Zou ¹, Li Fu Yang ^1,2, Lei Wang ¹, Su Sheng Yang ¹; Journal of Microbiology 2008;46(4):415-421.
DOI: https://doi.org/10.1007/s12275-008-0009-2
Published online: August 31, 2008

¹Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, and Key Laboratory of Agro-Microbial Resource and Application, Ministry of Agriculture, Beijing 100094, P. R. China, ²Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, and Key Laboratory of Physiology for Tropical Crops, Ministry of Agriculture, Danzhou Hainan 571737, P. R. China

Corresponding author: Su Sheng Yang , Tel: 86-10-6273-2173,

Received: 3 January 2008 • Accepted: 24 June 2008

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Abstract

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.

Keywords: Halobacillus aidingensis;Na+/H+ antiporter;nhaH;Inverse PCR

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Cloning and Characterization of a Na+/H+ Antiporter Gene of the Moderately Halophilic Bacterium Halobacillus aidingensis AD-6T

J. Microbiol. 2008;46(4):415-421. Published online August 31, 2008

DOI: https://doi.org/10.1007/s12275-008-0009-2

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