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The molecular cloning and structural analysis of a cytochrome P450 (CYP71D500) encoding gene from ajowan (Trachyspermum ammi L.) medicinal plant | ||
Iranian Journal of Genetics and Plant Breeding | ||
دوره 12، شماره 1 - شماره پیاپی 23، تیر 2023، صفحه 1-10 اصل مقاله (792.4 K) | ||
نوع مقاله: Research paper | ||
شناسه دیجیتال (DOI): 10.30479/ijgpb.2023.18141.1332 | ||
نویسندگان | ||
Shirin Jalal-Dowlatshahi1؛ Seyed Ahmad Sadat-Noori* 1؛ Seyed Mohammad Mahdi Mortazavian1؛ Mehdi Soltani Howyzeh2؛ Kasra Esfahani* 3 | ||
1Department of Agronomy and Plant Breeding Sciences, College of Aburaihan, University of Tehran, Tehran, Iran. | ||
2Department of Genetics and Plant Breeding, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran. | ||
3Department of Plant Bioproducts, Institute of Agricultural Biotechnology (IAB), National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran. | ||
تاریخ دریافت: 21 آذر 1401، تاریخ بازنگری: 15 مهر 1402، تاریخ پذیرش: 16 مهر 1402 | ||
چکیده | ||
Plant phenolic monoterpenes, such as thymol and carvacrol, have a wide range of applications in medicinal, pharmaceutical and other industries. Ajowan is an aromatic medicinal plant from the Apiaceae family with thymol as an active component of its seeds. The seeds of ajowan are valuable for medicinal purposes because their essential oil contains active substances of thymol, carvacrol, γ-terpinene, and p-cymene. Cytochrome P450 (CYP) –related genes have an indispensable role in the biosynthetic pathway of thymol and other substances in ajowan. A cytochrome P450 gene (CYP71D500) with a high expression level was isolated from the ajowan, cloned and sequenced. The sequencing information from the previous RNA-Seq study confirmed that the isolated gene belongs to the plant CYP71 clan, with a 1654 bp length containing two exons and a 115 bp intron. The full-length cDNA of CYP71D500 was also cloned. The sequencing of CYP71D500 cDNA showed the complete homology of CYP71D500 cDNA and exon regions of the CYP71D500 genomic sequence. The sequencing analysis of CYP71D500 cDNA also revealed a mutation that changed isoleucine to valine amino acid. The 3D structural analysis of the enzyme showed that the modified amino acid is not located in the enzyme’s active site. Therefore, this cannot probably affect the synthesis of thymol. The isolated gene and cDNA could be used for the metabolic engineering of ajowan and other medicinal plants with active phenolic monoterpenes. It is also applicable for identifying the different functions of the cloned CYP gene in ajowan or other medicinal plants. | ||
کلیدواژهها | ||
Ajowan؛ Apiaceae؛ Cytochrome P450؛ CYP71D500؛ Metabolic engineering؛ Monoterpenes | ||
عنوان مقاله [English] | ||
کلون سازی مولکولی و آنالیز ساختاری ژن کد کننده سیتوکروم P450 (CYP71D500) از گیاه دارویی زنیان (Trachyspermum ammi L.) | ||
نویسندگان [English] | ||
شیرین جلال دولتشاهی1؛ سید احمد سادات-نوری1؛ سید محمد مهدی مرتضویان1؛ مهدی سلطانی حویزه2؛ کسری اصفهانی3 | ||
1گروه زراعت و علوم اصلاح نباتات، پردیس ابوریحان، دانشگاه تهران، تهران، ایران. | ||
2گروه ژنتیک و اصلاح نباتات، واحد اهواز، دانشگاه آزاد اسلامی، اهواز، ایران. | ||
3پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری، تهران، ایران. | ||
چکیده [English] | ||
مونوترپن های فنلی گیاهی مانند تیمول و کارواکرول کاربردهای گسترده ای در پزشکی، صنایع دارویی و سایر صنایع دارند. زنیان گیاهی دارویی معطر از خانواده چتریان است که تیمول ماده موثره بذور آن است. دانههای زنیان برای اهداف دارویی ارزشمند هستند، زیرا اسانس آنها حاوی مواد فعال تیمول، کارواکرول، گاما-ترپینن و پارا-سایمن است. ژنهای سیتوکروم P450 (CYP) نقش مهمی در مسیر بیوسنتزی تیمول و سایر مواد در گیاه زنیان دارند. در این مطالعه یک ژن سیتوکروم P450 (CYP71D500) با سطح بیان بالا از گیاه دارویی زنیان جدا شده، شبیهسازی و توالییابی شد. اطلاعات توالی یابی از مطالعه RNA-Seq قبلی تایید کرد که ژن جدا شده متعلق به خانواده CYP71 گیاهی با طول 1654 جفت باز حاوی دو اگزون و یک اینترون 115 جفت باز است. cDNA کامل CYP71D500 نیز کلون شد. توالی یابی cDNA CYP71D500 همسانی کامل cDNA CYP71D500 و مناطق اگزونی آن را با توالی CYP71D500 ژنومی نشان داد. تجزیه و تحلیل توالی cDNA CYP71D500 جهشی را نشان داد که ایزولوسین را به اسید آمینه والین تغییر می دهد. تجزیه و تحلیل ساختاری سه بعدی آنزیم نشان داد که اسید آمینه تغییریافته در محل فعال آنزیم قرار ندارد. بنابراین احتمالا این جهش نمی تواند بر سنتز تیمول تأثیر بگذارد. ژن و cDNA جدا شده را می توان برای مهندسی متابولیک این گیاه و سایر گیاهان دارویی با مونوترپن های فنلی فعال استفاده کرد. همچنین برای شناسایی عملکردهای مختلف ژن CYP کلون شده در زنیان یا سایر گیاهان دارویی بکاربرد. | ||
کلیدواژهها [English] | ||
زنیان, اپیاسه, سیتوکروم پی450, CYP71D500, مهندسی متابولیت, منوترپنها | ||
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