1دانشجوی دکتری پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج
2دانشیار، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج
3دانشیار پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج
چکیده
در حال حاضر، انتقال ژرم لاین در حشرات روشی گسترده برای تجزیه و تحلیل عملکرد ژن و توسعۀ گونه‌های اصلاح‌شدۀ ژنتیکی است. اطلاعات زیادی دربارۀ نشو و نمای قسمت‌هایی از مغز جنین حشرات موجود نیست. به این منظور، چندین لاین تراریخت T. castaneum به هدف تعیین نقش برخی ژن‌های درگیر در نشو و نمای مغز ایجاد گردید. توالی کامل ژنوم T. castaneumمشخص شده است و روش‌هایی نیز برای ایجاد حشرات تراریخت این گونه ارائه شده است. در این پژوهش از عنصر متحرک Bacpiggy به عنوان ناقل بهT. castaneum استفاده شد. پلاسمید دهنده حاوی ناحیۀ ژنی مورد نظر، به جنین استرین چشم‌سفید (Tc vermilion-deficient) تزریق گردید. لاین جهش‌یافتۀ چشم‌سفید، دارای آسیب در ژن vermilionاست، اما از طرفی پلاسمید دهندۀ مورد نظر دارای ژن vermilionTc و تحت پروموتر 3xP3 است که سوسک‌های تراریخت را قادر به بیان ژن مذکور و در نتیجه ایجاد رنگ تیرۀ چشم می‌کند. در این مطالعه، لاین‌های تراریخت حاوی نواحی تنظیمی برخیژن‌های مهم در نشو و نمای مغز ایجاد شدند. این لاین‌ها به منظور ردیابی رشد نئورو بلاست‌ها از ابتدای رشد تا ساختار کاملاً رشدیافته در مغز حشرۀ تراریخت، ساخته شدند. به‌علاوه، این لاین‌ها امکان تشخیص ژن‌ها و مطالعات عملکردیشان را در مغز T. castaneum فراهم خواهند آورد.
Generation of transgenic lines for studying of the brain development in Tribolium castaneum (Coleoptera: Tenebrionidae)
نویسندگان [English]
Mahdiyeh Bigham1؛ Vahid Hosseininaveh2؛ Hossein Allahyari3
1Ph.D. Student , Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
2Associate Professor, Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
3Associate Professor, Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
چکیده [English]
Germ-line transformation of insects is now widely used for analyzing gene function and for the development of genetically modified strains for various purposes. There is not enough information on the embryonic development of some brain parts in insects. In order to study the genetic control of embryonic brain development, we established some transgenic lines to identify some genes involved in embryonic brain development. The genome has been sequenced and transgenic approaches are established for this model organism. Transposable element piggyBac was used as a vector for the transformation of T. castaneum. Tc-vermilion line- a Tribolium eye-color mutant- was also used for the microinjection of the donor vector. The donor vector contained the Tc-vermilion gene under the control of 3xP3 promoter, resulting in black eyes as a marker of their identity. In the present study, several transgenic lines bearing the regulatory regions of some importantgenes were generated. Additionally, transgenic lines for tracing of neuroblast development from the onset to the respective fully developed structure in the brain were established. Taken together, valuable tools which allow investigations of the complex genetic network needed for embryonic brain development in T. castaneum were established.Furthermore, the system will allow identification of genes and their functions in Tribolium brain formation.
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