نوع مقاله : مقاله پژوهشی
نویسندگان
1 گروه شیلات، دانشکده علوم وفنون دریایی، دانشگاه هرمزگان، بندرعباس، ایران.
2 گروه بیوتکنولوژی، زیست فناوری پزشکی و صنایع داروئی، سازمان تحقیقات علمی و صنعتی ایران، تهران، ایران.
3 گروه علوم و صنایع غذایی، دانشکده کشاورزی، دانشگاه صنعتی اصفهان، اصفهان، ایران.
4 گروه مهندسی ژنتیک - ژنومیکس، پژوهشکده بیوتکنولوژی کشاورزی ایران (ABRII) واحد اصفهان، اصفهان، ایران.
چکیده
در آبزی پروری مدرن باید مصرف انرژی، مواد خام مورد استفاده و اثرات زیست محیطی در نظر گرفته شوند لذا در طول دهه اخیر، آنزیم فیتاز توسط کارخانه های صنایع خوراک آبزیان به عنوان مکمل آنزیمی برای افزایش قابلیت هضم و جذب منابع پروتئینی گیاهی جیره، بهبود عملکرد رشد و کاهش آلودگی فسفر در محیط های آبی استفاده شده است. در این مطالعه با بهره گیری از سیستم تناوب‌هایِ کوتاهِ پالیندرومِ فاصله‌دارِ منظمِ خوشه‌ای (CRISPR: Clustered Regularly Interspaced Short Palindromic Repeats) اقدام به انتقال و درج ژن فیتاز باکتریایی به میکروجلبک مدل Chlamidomonas reinhardtii در جایگاه طراحی شده ژن نیترات ردوکتاز (NR) گردید و ویرایش صحیح ژنوم ریزجلبک با روش PCR بررسی و مورد تایید قرار گرفت. همچنین تایید کارآیی و بیان ژن فیتاز جایگذاری شده در جایگاه NR با استفاده از ژل SDS PAGE مورد بررسی قرار گرفت. نتایج کشت جلبک، حضور کلنی‌های مثبت دارای ویرایش صحیح بر روی محیط انتخابی در طول 15 نسل در دوره زمانی 150 روزه را نشان داد که خود نشان دهنده پایداری ویرایش صحیح بدون خاموشی ژن بود. همچنین ترجمه ژن فیتاز به عنوان تأیید بیان ژن انتقالی درج شده در سطح پروتئین توسط SDS PAGE بر روی پنج کلنی حاوی ویرایش صحیح و یک کلنی نمونه کنترل مورد تأیید قرار گرفت که نشان دهنده موفقیت رونویسی و ترجمه ژن فیتاز درج شده در جایگاه اگزون 2 ژن NR ریز جلبک است. با توجه به مزایای تولید و استفاده از آنزیم فیتاز نوترکیب در جیره های غذایی آبزیان از جمله استفاده بیشتر از منابع پروتئینی گیاهی، کاهش هزینه های تولید و کاهش غنی شدن بیش از حد مواد آلی در اکوسیستم های آبی، استفاده از این روش برای توسعه صنعت خوراک آبزیان و آبزی پروری مدرن توصیه می شود.
کلیدواژه‌ها
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