آنالیز پایداری بیان برخی از ژن‌های مرجع به منظور مطالعات بیان ژن در برنج با استفاده از Real-time PCR

نوع مقاله : مقاله پژوهشی

نویسندگان

1 عضو هیئت علمی دانشگاه صنعتی شاهرود

2 عضو هیات علمی دانشگاه صنعتی شاهرود

3 دانشجوی دکتری

چکیده

با ابداع واکنش زنجیره‌ای پلی‌مراز در زمان واقعی تحول عظیمی در زمینه تجزیه بیان ژن در موجودات زنده ایجاد شد، چراکه یکی از روش‌های مناسب برای ارزیابی میزان بیان ژن‌ها محسوب می‌گردد. در این روش، برای ارزیابی دقیق بیان ژن کنترل خطا بین نمونه‌ها ضروری می‌باشد. روشی که به صورت گسترده برای کنترل خطا مورد استفاده قرار می‌گیرد، نرمال کردن سطوح RNA با یک ژن مرجع یا خانه‌دار می‌باشد. این مطالعه به منظور بررسی کارایی10 ژن خانه‌دار در گیاه برنج در شرایط تنش شوری انجام شد. تیمار شوری صفر و 300 میلی‌مولار بعد از گذشت 8 هفته از رشد گیاهان اعمال گردید. همچنین در زمان‌های صفر، 6، 12، 24، 48 و 72 ساعت پس از اعمال تیمار شوری، از اندام‌های ریشه و برگ نمونه‌گیری صورت گرفت. نتایج حاصل از تجزیه و تحلیل داده‌ها با استفاده از نرم افزار geNorm نشان داد که ژن‌های eIF-4a و ACT7 در بافت ریشه و ژن‌های GAPDH و ACT11 در بافت برگ به عنوان پایدارترین ژن‌های مرجع می‌توانند در نرمال‌سازی مطالعات بیان ژن مورد استفاده قرار بگیرند. بر اساس آماره توصیفی در برنامه BestKeeper ژن eIF-4a در اندام ریشه و ژن‌های UBQ10 و eIF-4a در اندام برگ دارای بیشترین همبستگی با شاخص BestKeeper (به ترتیب 885/0، 891/0و 886/0) می‌باشد. در مجموع نتایج این تحقیق نشان داد که ژن‌های eIF-4a، ACT7، UBQ10 ، GAPDH و ACT11 می‌توانند به عنوان ژن‌های مرجع مناسب در گیاه برنج به منظور نرمال‌سازی داده‌های بیانی مورد استفاده قرار بگیرند.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Analysis of the stability of housekeeping genes expression for studying gene expression in rice using by Real-time PCR

نویسندگان [English]

  • Mohammad Reza Amerian 2
  • Arman Beyraghdar Kashkooli 3
2 academic staff
3 Ph.D student
چکیده [English]

A significant progress has been made in the expression profiling of living organism with invention of real time PCR, because this method is one of the suitable methods for evaluating the expression of genes. In this method, it is crucial to control the error observed between samples. Normalization with a house keeping gene is widely utilized to check the errors observed among samples in this method. In the current study, we evaluated expression patterns of 10 housekeeping genes under salinity stresses. We treated the eight- week old plants under salinity stress at concentrations of 0 and 300 Mm. Samples of leaves and roots were collected from the treated plants at 0, 12, 24, 48 and 72 hour post treatment. Expression analysis of the obtained data was performed via geNorm software and it was shown that the eIF-4a as well as ACT7 genes in the root tissues and GAPDH and ACT11 in the leaf tissues was constitutively expressed. Based on the results gained from the analysis trough Best Keeper, the eIF-4a gene (in root) and UBQ10 and eIF-4a (in leaf) showed the highest correlation with the index of Best keeper. Thus, we concluded that the eIF-4a, ACT7, GAPDH, UBQ10 and ACT11 genes could be considered as suitable housekeeping genes to be used in the normalization of the derived from Rice.

کلیدواژه‌ها [English]

  • Oryza sativa
  • Real time PCR
  • Salinity stress
  • Reference gene and BestKeeper
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