| As important transcription factors in the process of skeletal muscle formation,myogenic regulatory factors(MRFs)play a key role in regulating the differentiation and proliferation of skeletal muscle cells.MRFs family encodes 4 transcription factors,nemely Myf5,Myf6,MyoD and MyoG,which jointly control the formation and development of muscle and also have an important impact on meat flavor.In order to obtain the gene coding sequence(CDS)of MRFs in Sika deer,and explore the differential expression of this gene in muscle tissues of different parts of male and female Sika deer,myocardium,longissimus dorsi,quadriceps femoris,gluteus maximus and intercostal muscle tissues of adult healthy male and female Sika deer were selected as experimental materials.CDS of Myf5,Myf6,MyoD and MyoG genes were cloned with c DNA of gluteus maximus as template for sequencing and bioinformatics analysis.The relative m RNA expression of Myf5,Myf6,MyoD,MyoG genes in each muscle tissue was determined by real-time fluorescence quantitative PCR.The results of this study are as follows:1.CDS of Myf5,Myf6,MyoD,MyoG genes in Sika deer were cloned,the lengths of which were 768 bp,729bp,685 bp,684bp;after alignment in Gen Bank,it could be clustered into the same branch with CDS of domestic cattle(Gene ID: 281335),sheep(Gene ID:100188930),Cervus elaphus(Gene ID: 122693872)and Cervus canadensis canadensis(Gene ID: 122452506),with the closest genetic distance and homology of 99.61%,99.17%,100%and 100%,respectively.Myf5,Myf6,MyoD and MyoG all had strong hydrophilicity;the proportion of serine,serine,alanine and isoleucine were the highest in amino acid composition,accounting for 13.3%,10.7%,11.8% and 11.0% of the total amino acids;the secondary structure of the protein consists of α-helix,extended chain,random coil and β-folding.2.Myf5,Myf6,MyoD and MyoG genes of Sika deer were expressed in all muscle tissues of female and male Sika deer.Among them,the expression of Myf5 and MyoG genes in gluteus maximus tissue of male and female Sika deer and intercostal muscle tissue of female Sika deer were significantly higher than those in other tissues(P<0.05).The expression of Myf6 gene in quadriceps femoris of male and female Sika deer was significantly higher than that in other tissues(P<0.05);the expression level of MyoG gene in gluteus maximus of male and female Sika deer was significantly higher than that in other tissues(P<0.05).In addition,the expression levels of Myf5,Myf6,MyoD and MyoG genes in all muscle tissues except myocardium and intercostal muscles showed the same trend in male and female Sika deer,but there was no significant difference between males and females(P>0.05).In conclusion,Myf5,Myf6,MyoD,MyoG genes all play the role of myogenic regulation in gluteus maximus,longissimus dorsi,quadriceps femoris,myocardium and intercostal muscle tissues of female and male Sika deer,and gender has a significant effect on the role of MRFs in myocardium and intercostal muscles(P<0.05),but has no significant effect on the role of MRFs in gluteus maximus,longissimus dorsi and quadriceps femoris(P>0.05).It is meaningful for domestic Sika deer industry to grasp the differential expression of MRFs in different muscle tissues,so as to provide data basis for improving venison production performance and economic benefits of deer industry. |
