بیان موقت پروتئین فلورسنت سبز در تربچه (Raphanus sativus) با استفاده از وکتور بر پایة ویروس موزاییک شلغم

نوع مقاله : Research Paper

نویسندگان

گروه بیوتکنولوژی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان، ایران، کدپستی: 711-76315

چکیده

از گیاهان تراریخت می­توان به جای بیوراکتور برای تولید ارزان مواد شیمیایی و دارویی استفاده کرد. در سال­های اخیر، ناقلان ویروسی به طور گسترده در نقش ابزار بیوتکنولوژی موفق برای بیان طیفی گسترده از پروتئین­های نوترکیب در گیاهان توسعه­یافته­ عمل می­کنند. مزیت کاربرد ناقلان ویروسی به سبب نظام همانندسازی خودکار، افزایش سطح بیان ژن در زمانی کوتاه است. جُستار پیش­رو، نخستین مطالعة ناقل بر پایة TuMV در گیاه تربچه به شمار می­رود که از ناقل بر پایة ویروس موزاییک شلغم (TuMV)، برای تبیین موقت پروتئین فلورسنت سبز (GFP) در گیاه تربچه استفاده کرده­است. برگ­ها به صورت مکانیکی به کمک خراش با کربوراندوم تلقیح، پس از گذشت 14 روز برای ارزیابی برداشت شدند و انتقال و بیان پروتئین موردنظر با استفاده از میکروسکوپ فلورسنت، RT-PCR، SDS-PAGE و دات بلات تأیید گردید. آنالیز الایزا از بیان فراوانی پروتئین GFP در گیاهان تلقیح شده نسبت به فرم وحشی حکایت دارد. نتایج پژوهش نشان می­دهد که ناقل بر پایة TuMV کارایی بیشتری برای بیان پروتئین خارجی در گیاه تربچه دارد.

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