Vacuolar H+‐pyrophosphatase plays an important role in plant growth and stress response. By taking use of in-silico cloning and RT-PCR, here we isolated a vacuolar H+‐PPase cDNA (named LbVP1) from Lycium barbarum and analyzed the spatiotemporal expression pattern and its sequence structure. The open reading frame of LbVP1 expanded 2301 bp which encodes for 766 amino acid residues. Sequence alignment suggested a 87.9% identity of the putative LbVP1 protein in relation to Arabidopsis thaliana VP1. This deduced protein contained conserved domains and amino acid residues with the H+‐PPase proteins from other plants. By transient expression in Arabidopsis thaliana protoplast, LbVP1 was found mainly in the vacuolar membrane. The quantitative real-time PCR reflected variations on expression of LbVP1 varied in flower, leaves and fruit of different development stages. Under drought stress, its transcriptional level of this gene in leaves and fruits was significantly different in relative to the controls. Thus, these results will provide insights for elucidating the molecular mechanism of LbVP1 in response to the drought stress.