Cloning and sequence analysis of HKT1;4 gene fragment fromhalophyte Puccinellia tenuiflora

Abstract: In order to investigate Na+ exclusion mechanism of HKT1;4(HKT7) in Puccinellia tenuiflora, a pair of degenerate primers were designed based on the conserved sequences of the HKT1;4 genes registered in GenBank from other higher plants and reverse transcription was performed immediately after total RNA was extracted from the root. The partial cDNA fragment was amplified by PCR and the PCR product was subcloned into pMD19T Simple vector and sequenced after PCR detection. Sequencing results indicated that the length of the amplified cDNA fragment was 629 bp encoding 209 amino acid. Sequence analysis suggested that the nucleotide sequence and the translated amino acid sequence shared over 74% and 60% of homology to HKT1;4 gene sequences from other higher plants, respectively. It was confirmed that the fragment that we cloned belongs to the HKT1;4 gene family, therefore, we named it as PutHKT1;4.