Partial Cloning and DNA Sequencing of Terpene Synthase-4 Gene (TPS-4) in Origanum onites L.

Abstract

Thyme plant is commercially widely produced plant species that used in many fields as drug, cosmetic and spice. Especially Origanumonites L. plant species belonging to the Lamiaceae family is most cultivated thyme species in the world. Many thyme species contain plenty of volatile oil within. Terpenes are compounds that providing aromatic flavor to plants and the smell of their own. Terpenes constituted basis structure of volatile oil in aromatic plants were cultivated for years. Terpene synthases synthesize various terpenes in volatile oil produced as secondary metabolite. The aim of this study is to determine existence of terpene synthase 4 (TPS-4) gene in O. onites L. and to clone. The degenerative primers were designed for TPS-4 gene. RNA isolation from O. onites was performed with TRIzol reagent. cDNAs were obtained by revers transcription from mRNAs. The partial DNA fragments of TPS-4 gene were amplified by RT-PCR method using designed primers. Amplified TPS-4 gene fragments were added to pGEM-T Easy vector with ligation reaction. Recombinant vectors including TPS-4 gene fragments were transformed to XLI-Blue strain of E. coli. The amplified vectors in host bacteria were isolated for DNA sequencing. DNA sequence analysis was performed and DNA sequence was analyzed by using NCBI database. DNA sequence data were confirmed for TPS-4 gene. The sequence data presented in this study is the first report for a TPS-4 gene in O. onites. The DNA sequence and primers obtained from this study can be used as an important initial data in TPS-4 gene expression and biotechnology studies in the future.