بررسی بیان ژن MYH7 در بافت‌های ران، دست و راسته بره‌های پرواری نژاد کرمانی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد بخش علوم دامی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، ایران

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

3 استاد بخش علوم دامی، دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، ایران

4 استادیار، دانشکده کویرشناسی، دانشگاه سمنان، سمنان، ایران

چکیده

هدف: ماهیچه های اسکلتی حدود 40 درصد وزن بدن را تشکیل می‌دهند و عملکردهای زیادی، مانند حفظ انرژی مورد نیاز، حفظ وضعیت بدنی و محافظت از بافت‌های نرم را به عهده دارند. رشد طبیعی ماهیچه‌های اسکلتی پیش نیاز حیوانات برای حفظ فعالیت‌های طبیعی زندگی و متابولیسم است و هر گونه رشد غیر طبیعی ماهیچه‌ها منجر به بیماری می‌شود. بر اساس تجزیه و تحلیل پروفایل های بیان ژن در ماهیچه اسکلتی ژن MYH7 به عنوان ژن کاندیدای مربوط به انقباض عضلانی شناسایی شده است. هدف این مطالعه، بررسی بیان ژن MYH7 در بافت‌های ران، دست و راسته بره‌های پرواری نژاد کرمانی بود.
مواد و روش‌ها: در پژوهش حاضر نمونه برداری از شش راس بره نر کرمانی  از بافت‌های ران، دست و راسته با سه تکرار در هنگام کشتار انجام شد و  تعداد کل 54 نمونه‌ به دست آمد. RNA کل با استفاده از کیت استخراج (شرکت دنازیست) انجام شد.  کیفیت و کمیتRNA  استخراج شده با روش الکتروفورز روی ژل آگارز 2 درصد و با استفاده از دستگاه نانودراپ مورد ارزیابی قرار گرفت. برای سنتز  cDNAاز روی RNA استخراج شده از کیت سنتز cDNA شرکت parstous استفاده شد. از واکنش real time PCR به روش Syber Green برای بررسی میزان نسبی بیان ژن‌ها استفاده شد. از روش Pfaffl et al. (2002) برای تجزیه و تحلیل داده های حاصل از real time PCR استفاده شد.
نتایج: نتایج بررسی کیفیت RNA­های استخراج شده در طول موج A260/A280 نشان داد که کیفیت 8/1-9/1 مناسب و مطلوب است. به علاوه، مشاهده دو باند 18S و 28S برای RNA روی ژل آگارز نشان داد که RNA سالم است و نبود باند اضافی روی ژل نشان داد که RNA استخراج شده خالص است. نتایج منحنی­های real time PCR و الکتروفورز محصولات PCR روی ژل آگارز (2 درصد) نشان داد که ژن MYH7 در بافت­های دست، ران و راسته بره‌های نر کرمانی تکثیر و بیان می‌شود. در بافت‌های دست، ران و راسته باند 283 جفت بازی برای ژن MYH7 و برای ژن بتااکتین باند 112 جفت بازی در همه­ نمونه­ها مشاهده شد. این مشاهدات نشانگر این بود که آزمایش صحیح انجام شده است و قطعات مورد نظر به درستی تکثیر شده‌اند. نتایج حاصل از این پژوهش نشان داد که این ژن در سه بافت دست، ران و راسته بیان می‌شود.
نتیجه‌گیری: در پژوهش حاضر مشخص شد که ژن MYH7 در بافت‌های ماهیچه‌های ران، دست و راسته بیان می‌شود. لذا، می‌توان نتیجه گرفت که ژن MYH7 در ماهیچه‌های مختلف، که برای تولید بیشتر گوشت در نژادهای گوشتی بسیار ضروری هستند نقش ایفا می‌کند.

کلیدواژه‌ها

عنوان مقاله [English]

The expression of MYH7 gene in femur, humeral muscle and back muscle tissues of fattening lambs of the Kermani breed

نویسندگان [English]

  • Samira Shokri 1
  • Amin Khezri 2
  • Mohammadreza Mohammadabadi 3
  • Hamid Kheyrodin 4
1 MSc Student, Animal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
2 Associate Professor, Department of Animal Science, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.
3 Professor, Animal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
4 Assistant Professor, Semnan University, Semnan, Iran
چکیده [English]

Objective
Skeletal muscles make up about 40% of body weight and are responsible for many functions, such as maintaining required energy, maintaining body condition, and protecting tissues. Normal growth of skeletal muscles is a prerequisite for animals to maintain normal functions and metabolism, and any abnormal growth of muscles leads to disease. Based on the analysis of gene expression profiles in skeletal muscle, MYH7 gene has been identified as a candidate gene related to muscle contraction. The aim of this study was to investigate the expression of MYH7 gene in femur, humeral muscle and back muscle tissues of fattening lambs of Kermani breed.
Materials and Methods
In this research, six Kermani male lambs were sampled from femur, humeral muscle and back muscle tissues at the time of slaughter. The total number of samples was equal to 54 samples (6 animals, 3 tissues and 3 repetitions of each tissue). Total RNA was extracted using a standard extraction kit. The quality and quantity of extracted RNA was evaluated using electrophoresis on 2% agarose gel and using nanodrop device. Parstous cDNA synthesis kit was used for the cDNA synthesis from the extracted RNA. Syber Green real time PCR reaction was used to check the relative level of gene expression. The method of Pfaffl et al. (2002) was used to analyze the data obtained from real time PCR.
Results
The results of RNAs extracted at A260/A280 wavelength showed that the quality is suitable and desirable (1.9-1.8). In addition, the observation of two bands of 18S and 28S for RNA on agarose gel indicated that the RNA was intact and the absence of additional bands on the gel indicated that the extracted RNA was pure. The results of real time PCR curves and electrophoresis of PCR products on agarose gel (2%) showed that the MYH7 gene is amplified and expressed in the femur, humeral muscle and back muscle tissues of Kermani male lambs. For the studied tissues (femur, humeral muscle and back muscle), a band of 283 bp was observed for the MYH7 gene and a band of 112 bp was observed for the beta-actin gene in all samples. These observations indicated that the correct experiment was performed and the desired fragments were amplified correctly. The results of this research showed that this gene is expressed in the femur, humeral muscle and back muscle tissues.
Conclusion
In the present study, it was found that the MYH7 gene is expressed in the femur, humeral muscle and back muscle. Therefore, it can be concluded that MYH7 gene plays a role in different muscles, which are very necessary for more meat production in the sheep breeds

کلیدواژه‌ها [English]

  • MYH7 gene
  • femur
  • humeral muscle
  • back muscle

مراجع

جعفری احمدآبادی سید علی اصغر، عسکری­همت حشمت­اله، محمدآبادی محمدرضا (1402) تاثیر شاهدانه بر بیان ژن DLK1 در بافت‌ قلب بره‌های کرمانی. مجله بیوتکنولوژی کشاورزی، 15(1)، 217-234.
عرب پور رق آبادی زهرا، محمدآبادی محمدرضا، خضری امین (1400) الگوی بیانی ژن p32 در بافت‌های ران، دست، راسته و چربی پشت بره کرمانی. مجله بیوتکنولوژی کشاورزی، 13(4)، 183-200.
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، خیرالدین حمید، لطیفی آیگین، بابنکو اولنا ایوانیونا (1401) پروفایل بیانی ژن  DNAH1در بافت بیضه بز کرکی راینی. مجله بیوتکنولوژی کشاورزی، 14(3)، 243-256.
محمدآبادی محمدرضا، سفلایی محمد (1399). پروفایل بیانی mRNA مختص بافت ژن BMP15 در بز. مجله بیوتکنولوژی کشاورزی 12(3)، 208-191.
محمدی فر آمنه، محمدآبادی محمدرضا (1390) کاربرد نشانگرهای ریزماهواره برای مطالعه ژنوم گوسفند کرمانی. مجله علوم دامی ایران 42(4)، 344-337.
محمودی مریم ، آیت اللهی مهرجردی احمد ، محمدآبادی محمدرضا (1396) بررسی اگزون چهارم ژن کاپاکازئین گوسفند کرمانی با تکنیک PCR-RFLP. مجله بیوتکنولوژی کشاورزی، 9(3)، 128-119.
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مجله بیوتکنولوژی کشاورزی
دوره 15، شماره 2
خرداد 1402
صفحه 217-236

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