مجله بیوتکنولوژی کشاورزی (May 2023)
The expression of MYH7 gene in femur, humeral muscle and back muscle tissues of fattening lambs of the Kermani breed
Abstract
ObjectiveSkeletal 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 MethodsIn 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.ResultsThe 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.ConclusionIn 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
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