Endemik Sütçüler Kekiğinde (Origanum minutiflorum) Uçucu Yağ Sentezinde Rol Oynayan Bazı Genlerin İzolasyonu ve Karakterizasyonu

Öz

Origanum minutiflorum, Türkiye'de Isparta ilinin Sütçüler ilçesinde yetişen endemik bir türdür. Bu çalışmada O. minutiflorum bitkisinden uçucu yağ sentezinde görev alan 1-deoksi-D-ksilüloz fosfat redüktoizomeraz (OmDXR) ve terpen sentaz 1 (OmTPS1) genlerinin izolasyonu ve karakterizasyonu amaçlanmıştır. Bu amaç doğrultusunda kekik yapraklarından toplam RNA izole edilmiş ve hedef genlere özgü primerler ile genlerin sekans bilgileri elde edilmiştir. Sekans bilgileri BLAST programları kullanılarak, NCBI veri bankasında bulunan ve farklı türlerden elde edilen gen sekansları ile karşılaştırılmıştır. Nükleotid dizilerin protein dizisine dönüştürülmesinde ExPASy Tools, kodlanan proteinlerin 3 boyutlu yapısının oluşturulmasında 3D SWISS MODEL, nükleotid ve protein dizilerinin filogenetik ilişkileri, MEGAX 11 paket programı kullanılarak oluşturulmuştur. Çalışma sonucunda OmDXR genine ait tam uzunluktaki cDNA nükleotid uzunluğunun 1 700 baz çifti ve gene ait nükleotid dizisinin önceki çalışmalarda farklı türlerden elde edilen nükleotid dizileri ile %74.22-81.07, protein dizisinin ise %70.92-72.95 arasında benzerlik gösterdiği belirlenmiştir. OmTPS1 genine ait tam uzunluktaki cDNA nükleotid uzunluğunun ise 1 836 baz çifti ve gene ait nükleotid dizisinin önceki çalışmalarda farklı türlerden elde edilen nükleotid dizileri ile %75.99-89.58, protein dizisinin ise %51.66-69.88 arasında benzerlik olduğu tespit edilmiştir. Söz konusu çalışma O. minutiflorum türünde yapılan ilk moleküler temelli çalışma olmuştur. Dolayısıyla konu ile ilgili gelecekte yapılacak olan çalışmalar için bir kaynak özelliği taşıyacaktır.

Anahtar Kelimeler

• DXR
• Gen klonlama
• Origanum minutiflorum
• Terpen sentaz
• Uçucu yağ

Isolation and Characterization of Some Genes Involved in Essential Oil Synthesis in Endemic Sütçüler Thyme (Origanum minutiflorum)

Abstract

Origanum minutiflorum, is an endemic species found in the Sütçüler district of Isparta province, Turkey. This study aimed to isolate and characterize the 1-deoxy-D-xylulose-5-phosphate reductoisomerase (OmDXR) and terpene synthase 1 (OmTPS-1) genes, which play roles in essential oil biosynthesis in O. minutiflorum. For this purpose, total RNA was isolated from thyme leaves, and the sequence information of the target genes was obtained using gene-specific primers. The sequences were compared with those from other species in the NCBI database using BLAST programs. The nucleotide sequences were translated into protein sequences using ExPASy Tools, the three-dimensional structures of the encoded proteins were constructed with 3D SWISS-MODEL, and phylogenetic relationships of nucleotide and protein sequences were analyzed using the MEGA X version 11 software package. The results showed that the full-length cDNA nucleotide sequence of the OmDXR gene is 1,700 base pairs (bp), and its nucleotide sequence displayed 74.22%-81.07% similarity to nucleotide sequences from other species in previous studies, while the protein sequence showed 70.92%-72.95% similarity. The full-length cDNA nucleotide sequence of the OmTPS-1 gene was determined to be 1.836 bp, and its nucleotide sequence exhibited 75.99%-89.58% similarity to nucleotide sequences from other species, while the protein sequence showed 51.66%-69.88% similarity. This study represents the first molecular-based research on the O. minutiflorum species. Therefore, it will provide a valuable resource for future studies.

Keywords

• DXR
• Essential oil
• Gene cloning
• Origanum minutiflorum
• Terpene synthase

Kaynakça

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