Molecular cloning and bioinformatics analysis of a vacuolar Na+/H+ antiporter gene in Elytrigia intermedia

Abstract

TiNHX1, homologous with the TaNHX1 gene encoding a vacuole Na+/H+ antiporter was cloned from Elytrigia intermedia by RT-PCR, with primers designed according to the sequence of TaNHX1. The largest open reading frame of TiNHX1 gene has 1 641bp in length and encoded a protein of 546 amino acid residues. The estimated molecular weight and isoelectric points of the putative protein was 59.8 kDa and 8.0, respectively. Components of amino acids encoded by TiNHX1 contained 38 basic amino acids, 36 acidic amino acid, 256 hydrophobic amino acids and 129 polar amino acids. The predicted secondary structure composition for the protein has about 44% alpha helixes, 21% extended strand, 4% beta turn and 29% random coil. Hydrophobic analysis indicated that the TiNHX1 contained 10 potential transmembrane segments. Blast result and the phylogenetic analysis showed that TiNHX1, AtNHX1, OsNHX1, GmNHX1, TaNHX1 share a cluster.TiNHX1, homologous with the TaNHX1 gene encoding a vacuole Na+/H+ antiporter was cloned from Elytrigia intermedia by RT-PCR, with primers designed according to the sequence of TaNHX1. The largest open reading frame of TiNHX1 gene has 1 641bp in length and encoded a protein of 546 amino acid residues. The estimated molecular weight and isoelectric points of the putative protein was 59.8 kDa and 8.0, respectively. Components of amino acids encoded by TiNHX1 contained 38 basic amino acids, 36 acidic amino acid, 256 hydrophobic amino acids and 129 polar amino acids. The predicted secondary structure composition for the protein has about 44% alpha helixes, 21% extended strand, 4% beta turn and 29% random coil. Hydrophobic analysis indicated that the TiNHX1 contained 10 potential transmembrane segments. Blast result and the phylogenetic analysis showed that TiNHX1, AtNHX1, OsNHX1, GmNHX1, TaNHX1 share a cluster.