Journal of Cell & Tissue, Winter 2013, 3(4) Journal of Cell & Tissue (JCT) Research Article Winter 2013; 3(4): 307-318 Cloning and Study the Bioinfomatic Trait of Tropinone Reductase-II (TR II) Gene from Hyoscyamus niger Parsa M. M.Sc. 1, 2 , Garoosi G.A. Ph.D. 1* , Haddad R. Ph.D. 1 1. Agricultural Biotechnology Department, Faculty of Engineering and Technology, Imam khomeini international university, Qazvin, Iran. P.BOX: 34149-161818 2. Plant Ecology and Systematic Department, Applied Science Institute, Shaid Beheshti University, Tehran, Iran. P.BOX: 196151171 * Email corresponding author: a-garoosi90@yahoo.com Received: 24 Jun. 2012 Accepted: 22 Jan. 2013 Abstract Aim: The purpose of present research was extraction and cloning of tropinone reductase-II gene (tr-II) at antisense direction in pBI121 binary vector to provide transgenic plants with low rate of tropinone reductase-II enzyme and high production of scopolamine and hyoscyamine for future projects. Materials and methods: Total RNA was extracted from Iranian native Hyoscyamus niger roots, 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. Results: 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. Conclusion: 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. Keywords: Agrobacterium, Antisense, Bioinformatics, Cloning, Hyoscyamus niger, Tropinone reductase II, TR-II