Abstract

Origanum onites L. (oregano) is an aromatic plant used in medicine, cosmetics and different industrial fields. It includes essential oil and is extensively cultivated as aromatic plant in the world. Therefore, it is also preferred as a spice in foods. Terpene synthase-6 (TPS-6) is an enzyme responsible for synthesis of sesqui-terpenes in aromatic plants. The investigation of gene sequence information and expression characteristics of this enzyme in O. onites may contribute to increase of synthesis of terpenes as raw materials in medicine and cosmetic etc. industry. In this study, partial cloning and sequencing of the TPS-6 gene in the O. onites was aimed. Total RNA isolation from O. onites was performed with TRI Reagent solution. The mRNAs were transformed to cDNAs by Revers Transcription Method. The middle region of TPS-6 gene was amplified by PCR method using degenerative primers designed for TPS-6. Amplified DNA fragments were obtained at 481bp. size and ligated to PGEM T Easy plasmid vector with T4 DNA ligase. The recombinant plasmid vector was transferred to the XL1- Blue host cells (E. coli) and the cloned DNA was sequenced. The partial TPS-6 DNA sequence was confirmed in NCBI GenBank database and registered with an accession number (MF983853). The obtained DNA sequence was analyzed with InterProScan and Clustal Omega software in EBI database. The analysis results showed that this DNA sequence has a specific motif (DXXD) for TPS-6 catalyzing cyclization reactions. The phylogenetic dendrogram was drawn for comparison with other plant TPS-6 genes. O. onites TPS-6 sequence was determined to be very close to O. vulgare TPS-6 on the dendrogram than other plant TPS-6s. The obtained gene sequence information will guide biotechnological researches in the productivity increase of this gene using expression analysis.