Cloning and Characterization of a Sulfate Transporter Gene from Pimpinella brachycarpa

To investigate the regulation of sulfur uptake and subsequent sulfur metabolism in higher plants, sulfate transporter gene was identified and analyzed in Pimpinella brachycarpa. Total RNA from P. brachycarpa leaves derived from somatic embryos, was quantified and determined the quality by visualizing two distinct cytosolic rRNAs. The sulfate transporter gene using the biotin labeled Hvst 1 probe was under high induction and mRNA transcript of the gene was present in the cytosol. In the colony lift hybridization colonies hybridized with the probe might have recognized an insert sequence within the E. coli genome. Two positive clones were isolated from 3.6 x 105 amplified plaques. The isolated clone (PST829) contained an open reading frame encoding a polypeptide of 717 amino acids. Northern blot analysis indicated the constitutive accumulation of a 2.6 kb length tranacript in leaves and roots of 3-week-old seedlings.