الگوی بیانی ژن p32 در بافت‌های ران، دست، راسته و چربی پشت بره کرمانی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 بخش علوم دامی، دانشگاه شهید باهنر کرمان

2 بخش علوم دامی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان

3 دانشیار، گروه علوم دامی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، کرمان، ایران

چکیده

هدف: مطالعات نشان داده‌اند که p32 در بافت‌های فعال متابولیکی و به سرعت در حال رشد، مانند ماهیچه‌ها، تومورهای سینه، اپیدرم و تخمدان به شدت بیان می‌شود. یکی از اقدامات اساسی در حیوانات اهلی مطالعه ژن­ها و پروتئین­های مرتبط با صفات اقتصادی و مطالعه آنها در سطح سلولی یا کروموزومی است. هدف این مطالعه، بررسی الگوی بیانی ژن p32 در بافت‌های ران، دست، راسته و چربی پشت کمر بره کرمانی بود.
مواد و روش‌ها: نمونه‌برداری از بافت‌های ران، دست، راسته و چربی پشت کمر بره‌های نر کرمانی (12 نمونه از 3 حیوان و از هر بافت 3 تکرار) انجام شد. پس از انجماد سریع نمونه ها در داخل فریزر 80- نگهداری شدند. RNA کل با استفاده از کیت استاندارد استخراج شد. کیفیت و کمیتRNA  استخراج شده با روش الکتروفورز روی ژل آگارز و با استفاده از دستگاه نانودراپ مورد ارزیابی قرار گرفت. برای سنتز  cDNAاز کیت سنتز cDNA شرکت parstous استفاده شد. برای بررسی میزان نسبی بیان ژن‌ها از واکنش real time PCR به روش Syber Green استفاده شد. برای تجزیه و تحلیل داده های حاصل از real time PCR از روش Pfaffl et al. استفاده شد.
نتایج: بررسی کیفیت RNA­های استخراج شده نشان داد که کیفیت مناسب و مطلوب است. مشاهده دو باند 18S و 28S در RNA نشان دهنده سالم بودن آن و عدم وجود باند اضافی نشان دهنده خلوص آن بود. الکتروفورز محصولات PCR روی ژل آگارز و نتایج منحنی­های real time PCR نشان داد که ژن p32 در بافت­های ران، راسته و چربی پشت کمر بره‌های نر کرمانی بیان نشده است و فقط در بافت دست این حیوانات تکثیر شده است. مشاهده تک باند در محدوده bp281 برای ژن p32 در بافت­ دست و وجود باند در محدوده bp112 برای ژن بتااکتین در همه­ نمونه­ها، بیانگر صحت انجام آزمایش و تکثیر قطعه مورد نظر بود.
نتیجه‌گیری: با توجه به این که در این مطالعه بیان ژن p32 در گوسفند کرمانی در سن 11 ماهگی مطالعه شد و بیان این ژن بیشتر در دوران جنینی گزارش شده است، لذا عدم مشاهده بیان این ژن در بافت‌های ران، راسته و چربی پشت و بیان پایین آن در بافت دست تایید کننده نتایج مطالعات قبلی است. بیان کم این ژن در عضله دست می‌تواند حاکی از این باشد که این عضله هنوز در این سن دارای رشد است و یا ممکن است این بافت سرطانی شده باشد، چرا که این ژن بیشتر در بافت‌هایی بیان می‌شود که در حال رشد سریع هستند و یا درگیر سرطان هستند.

کلیدواژه‌ها


عنوان مقاله [English]

The expression pattern of p32 gene in femur, humeral muscle, back muscle and back fat tissues of Kermani lambs

نویسندگان [English]

  • zahra arabpour 1
  • Mohammadreza Mohammadabadi 2
  • Amin Khezri 3
1 : MSc Student, Animal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
2 Department of Animal Science, Faculty of Agriculture, College of Agriculture, Shahid Bahonar University of Kerman
3 Associate Professor, Department of Animal Science, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.
چکیده [English]

Objective
Studies have shown that p32 is highly expressed in metabolically active and rapidly growing tissues, such as muscles, breast tumors, epidermis, and ovaries. One of the basic measures in domestic animals is the study of genes and proteins related to economic traits and their study at the cellular or chromosomal level. The aim of this study was to investigate the expression pattern of p32 gene in femur, humeral muscle, back muscle and back fat tissues of Kermani lambs.
 
Materials and Methods
Samples of femur, humeral muscle, back muscle and back fat tissues of Kermani lambs were collected (12 samples from 3 animals and 3 replicates from each tissue). After rapid freezing, the samples were stored in a -80 freezer. Total RNA was extracted using a standard kit. The quality and quantity of the extracted RNA were evaluated by agarose gel electrophoresis and using a nanodrop device. For cDNA synthesis, parstous cDNA synthesis kit was used. Real time PCR reaction by Syber Green method was used to evaluate the relative expression of genes. To analyze the data obtained from real time PCR, the Pfaffl method was used.
 
 
Results
The quality of the extracted RNAs showed that the quality is appropriate and desirable. Observation of 18S and 28S bands in RNA indicated that it was intact and that no additional bands indicated its purity. Electrophoresis of PCR products on agarose gel and the results of real time PCR curves showed that the p32 gene was not expressed in femur, back muscle and back fat tissues of Kermani male lambs and was amplified only in the humeral muscle tissue of these animals. The observation of a single band in the range of 281 bp for the p32 gene in the humeral muscle tissue and the presence of a band in the range of 112 bp for the beta actin gene in all samples indicated the accuracy of the experiment and the amplification of the fragment.
 
Conclusion
Considering that in this study the expression of p32 gene in Kermani sheep at the age of 11 months was studied and the expression of this gene has been reported more in the embryonic period, so no expression of this gene was observed in femur, back muscle and back fat and its low expression in the humeral muscle tissue confirms the results of previous studies. Low expression of this gene in the humeral muscle tissue may indicate that the muscle is still growing at this age or that the tissue may be cancerous, as this gene is more expressed in tissues that are growing rapidly or are involved in cancer.

کلیدواژه‌ها [English]

  • Expression pattern
  • p32 gene
  • Tissue
  • Kermani sheep
توحیدی نژاد فاطمه، محمدآبادی محمدرضا، اسمعیلی زاده کشکوئیه علی، نجمی نوری عذرا (1393). مقایسه سطوح مختلف بیان ژنRheb  در بافت های مختلف بز کرکی راینی. مجله بیوتکنولوژی کشاورزی 6(4)، 50-35.
جعفری دره در امیر حسین، محمدآبادی محمدرضا، اسمعیلی زاده کشکوئیه علی، ریاحی مدوار علی (1395) بررسی بیان ژن CIB4  در بافت­های مختلف گوسفند کرمانی با استفاده از real time qPCR. مجله پژوهش در نشخوارکنندگان 4(4)، 132-119.
محمدآبادی محمدرضا (1399) بیان ژن ESR1 در بز کرکی راینی با استفاده از real time PCR‎. مجله بیوتکنولوژی کشاورزی 12(1)، 192-177.
محمدآبادی محمدرضا، سفلایی محمد (1399). پروفایل بیانی mRNA مختص بافت ژن BMP15 در بز. مجله بیوتکنولوژی کشاورزی 12(3)، 208-191.
محمدآبادی محمدرضا، کرد محبوبه، نظری محمود (1397) مطالعه بیان ژن لپتین در بافت‌های مختلف گوسفند کرمانی با استفاده از real time PCR. مجله بیوتکنولوژی کشاورزی 10(3)، 122-111.
محمدی فر آمنه، محمدآبادی محمدرضا (1390) کاربرد نشانگرهای ریزماهواره برای مطالعه ژنوم گوسفند کرمانی. مجله علوم دامی ایران 42(4)، 344-337.
واجدابراهیمی محمدتقی، محمدآبادی محمدرضا، اسماعیلی زاده علی (1394) بررسی تنوع ژنتیکی پنج جمعیت گوسفند ایرانی با استفاده از نشانگرهای ریزماهواره­ای. مجله بیوتکنولوژی کشاورزی 7(4)، 158-143.
 
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