همسانه‌سازی و بررسی پروموتر دائمی یک ژن پلی‌یوبی‌کوئیتین از گیاه نخود

نوع مقاله: علمی پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه اصلاح نباتات و بیوتکنولوژی، دانشگاه شاهد، تهران، ایران

2 استادیار، گروه پژوهشی مهندسی ژنتیک و ایمنی زیستی، پژوهشگاه بیوتکنولوژی کشاورزی ایران، سازمان تحقیقات آموزش و ترویج کشاورزی، کرج، ایران

چکیده

ایجاد گیاهان تراریخته نیازمند پروموترهای جدید است. با ظهور فناوری‌های توالی‌یابی نسل نو و تولید انبوه داده‌های ژنومی برای گونه‌های مختلف گیاهان زراعی که با توسعه ابزارهای بیوانفورماتیکی همراه شده‌، فرصت مناسبی برای شناسایی، جداسازی و بررسی ویژگی‌های پروموترهای جدید فراهم آمده است. به‌دلیل وجود ملاحظاتی در مورد استفاده از پروموترهای ویروسی در گیاهان تراریخته لزوم همسانه‌سازی و استفاده از پروموترهایی با منشأ گیاهی احساس می‌شود. پروموترهای دائمی با منشأ گیاهی معمولاً دارای عناصر تنظیمی غیر اختصاصی بوده و به سادگی قابلیت بیشتری را در به‌کارگیری ماشین رونویسی سلول گیاهی برای رونویسی از ژن‌ها دارا هستند. بر اساس نحوه بیان ژن‌ها، پروموترها به سه دسته دائمی، القایی و ویژه بافت طبقه‌بندی می‌شوند. ژن‌های خانه‌دار بهترین و مهمترین منبع برای جداسازی پروموترهای دائمی هستند. در این پژوهش، بیان ژن گزارشگر بتا-گلوکورونیداز تحت کنترل پروموتر ژن پلی‌یوبی‌کوئیتین 10 نخود (Cicer ariethinum) در بافت‌های توتون بررسی شد. عملکرد CaUBQ10 در برگ‌، ساقه و ریشه‌های گیاهان تراریخته پایدار توتون تأیید شد که نشان می‌دهد CaUBQ10 یک پروموتر دائمی است و می‌تواند انتخاب مناسبی برای بیان بالای ژن‌های مورد نظر در کلیه مراحل زندگی گیاه باشد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Cloning and characterization of a constitutive promoter of polyubiquitin gene from Cicer ariethinum

نویسندگان [English]

  • Niloufar Peykari 1
  • Katayoun Zamani 2
1 MSc student, Department of Agricultural Biotechnology, Shahed University, Tehran, Iran
2 Assistant professor, Department of Genetic Engineering and Biosafety, Agricultural Biotechnology Research Institute of Iran, Agricultural Research, Education and Extension Organization, Karaj, Tehran, Iran.
چکیده [English]

Producing transgenic plants need new promoters. Due to the advent of next generation sequencing technologies and the production of massive genomic data for various crop plant species paralleled by the development of bioinformatics tools, there is an opportunity ‎to identify, isolate and characterize new promoters. Because of public concern about using viral promoters, there is a need for cloning and using plant promoters in transgenic food crops. Native plant constitutive promoters may be composed of non-specific elements that are simply more efficient at protein recruitment for transcription. Promoters are classified as inducible, constitutive and tissue specific according to the nature of gene expression they regulate. Housekeeping genes are the best and most important sources for isolating the constitutive promoters. In this study, -Glucoronidase reporter gene expression mediated by a polyubiquitin promoter (CaUBQ10) from chickpea (Cicer ariethinum) was analyzed in tobacco tissues. The functionality of the CaUBQ10 was confirmed in leaves, stems, and roots of stably transformed tobacco plants and suggested that the CaUBQ10 is a constitutive promoter and may provide a valuable choice for high-level expression of target genes during the life cycle of a plant.

کلیدواژه‌ها [English]

  • CaMV35S
  • polyubiquitin
  • Cicer ariethinum
  • housekeeping gene
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