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

ObjectiveStudies 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 MethodsSamples 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.  ResultsThe 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. ConclusionConsidering 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.