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Screening a novel Na⁺/H⁺ antiporter gene from a metagenomic library of halophiles colonizing in the Dagong Ancient Brine Well in China
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Screening a novel Na⁺/H⁺ antiporter gene from a metagenomic library of halophiles colonizing in the Dagong Ancient Brine Well in China
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Screening a novel Na⁺/H⁺ antiporter gene from a metagenomic library of halophiles colonizing in the Dagong Ancient Brine Well in China
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Journal Article
Screening a novel Na⁺/H⁺ antiporter gene from a metagenomic library of halophiles colonizing in the Dagong Ancient Brine Well in China
2010
Overview
Metagenomic DNA libraries constructed from the Dagong Ancient Brine Well were screened for genes with Na⁺/H⁺ antiporter activity on the antiporter-deficient Escherichia coli KNabc strain. One clone with a stable Na⁺-resistant phenotype was obtained and its Na⁺/H⁺ antiporter gene was sequenced and designated as m-nha. The deduced amino acid sequence of M-Nha protein consists of 523 residues with a calculated molecular weight of 58 147 Da and a pI of 5.50, which is homologous with NhaH from Halobacillus dabanensis D-8T (92%) and Halobacillus aidingensis AD-6T (86%), and with Nhe2 from Bacillus sp. NRRL B-14911 (64%). It had a hydropathy profile with 10 putative transmembrane domains and a long carboxyl terminal hydrophilic tail of 140 amino acid residues, similar to Nhap from Synechocystis sp. and Aphanothece halophytica, as well as NhaG from Bacillus subtilis. The m-nha gene in the antiporter-negative mutant E. coli KNabc conferred resistance to Na⁺ and the ability to grow under alkaline conditions. The difference in amino acid sequence and the putative secondary structure suggested that the m-nha isolated from the Dagong Ancient Brine Well in this study was a novel Na⁺/H⁺ antiporter gene.
Publisher
Oxford, UK : Blackwell Publishing Ltd,Blackwell Publishing Ltd,Wiley-Blackwell,Oxford University Press
Subject
/ Bacillus
/ Biological and medical sciences
/ Brines
/ China
/ DNA
/ Domains
/ E coli
/ Escherichia coli - metabolism
/ Fundamental and applied biological sciences. Psychology
/ genes
/ Genetic Complementation Test
/ Homology
/ Hydrogen
/ mutants
/ Protein Structure, Secondary
/ Residues
/ Salts
/ Sequence Homology, Amino Acid
/ sodium
/ Sodium Chloride - metabolism
/ Sodium-Hydrogen Exchangers - chemistry
/ Sodium-Hydrogen Exchangers - genetics
