Cloning and bioinformatics analysis of ACPase from tung tree

ObjectiveThe full-length sequence of acid phosphatase(ACPase)gene in tung tree was cloned to provide ref-erences for further studies on the functions of ACPase. MethodThe primers were designed according to the full-length se-quence of acid phosphatase gene in transcriptome of tung tree. ACPase gene from tung tree seeds was cloned by RT-PCR. Bioinformatic analysis was used to predict cDNA sequence and its deduced nucleotide sequence similarity, physicochemical properties, hydrophobicity, transmembrane structure, secondary structure, and tertiary structure of ACPase. Multiple se-quences alignment and phylogenetic trees were also constructed. ResultACPase in tung tree was cloned. The largest open reading frame of ACPase had 1152 bp in length and encoded a polypeptide of 383 amino acid residues. The estimated molecu-lar weight and isoelectric point of the putative protein was 40.94 kDa and 5.30, respectively. The ACPase was a soluble unsta-ble protein. BLASTs search result and phylogenetic tree analysis showed that peptide encoded by ACPase of tung tree had 84%similarity with ACPase of Populus trichocarpa. ACPase of tung tree had conservative regions of the phosphatase gene family. Most amino acid encoded by ACPase belonged to hydrophilic amino acid, and its protein secondary structure was mainly con-sisted of irregular coli,α-helix andβ-sheet. ACPase protein contained one transmembrane spiral region with its amino acid in 136-156 and one suspected transmembrane region with its amino acid in 255-275. Every transmembrane region was spirals consisted of 20 amino acid residues. N-terminal domain and C-terminal domain of the transmembrane protein were on one side of cytomembrane cytoplasm.ConclusionThe cloned ACPase of tung tree may be involved in the process of phosphorus nutri-tion in plants and the decomposition of soil organic phosphorus.