Cloning and Study the BioinfomaticTrait of TropinoneReductase-II (TR II) Gene fromHyoscyamusniger

The purpose of present research was extraction and cloning of tropinonereductase-II gene (tr-II) at antisense direction in pBI121 binary vector to provide transgenic plants with low rate of tropinonereductase-II enzyme and high production of scopolamine and hyoscyamine for future projects. Total RNA was extracted from Iranian native Hyoscyamusnigerroots, and the interest gene after cDNA synthesis and cloning at antisense direction in pBI121 binary vector, was transfered to Agrobacterium tumefacience. Accurate cloning was studied through 3 methods; enzymatic digestion, PCR and DNA sequencing. The bioinformatic characters of the gene were then surveyed. Three used methods confirmed true cloning in high efficiency. Nucleotide sequence of the gene revealed the 783 bp in length, encoding a polypeptide of 260 amino acid residues, with high similarity to that one registered in NCBI. The predicted molecular mass and isoelectric point of deduced polypeptide were 28437.3 Da and 5.46, respectively. Protein structures were not completely similar to those previously reported at PDB data base. Also, phylogenic study demonstrated that this gene belongs to the group I of TRs. Due to successful cloning and high similarity of nucleotide and polypeptide sequences of gene with those recorded in world data bases; it is expected to get success in access to main purpose.