Cloning,Bioinformatics Analysis and Tissue Expression of MYL9 Gene in Meiren Yak
[Objective]The objective of this experiment was to clone the CDS region of myosin light chain 9(MYL9)gene in Meiren yak and perform bioinformatics analysis,and detect the tissue expression characteristics of MYL9 gene,so as to provide a theoretical basis for exploring the function of this gene.[Method]Using the cDNA of Meiren yak muscle tissue as template,PCR was used to amplify and clone the CDS region sequence of Meiren yak,similarity comparison with other species and phylogenetic tree construction were carried out.Bioinformatic analysis of MYL9 protein was performed by online software.The expression of MYL9 gene in heart,liver,spleen,lung,kidney,muscle,fat and testis of Meilen yak was detected by Real-time quantitative PCR.[Result]The CDS region of MYL9 gene in Meiren yak was 516 bp,encoding 171 amino acids.Phylogenetic tree results showed that Meiren yak had the closest genetic relationship with Bos mutus and Bos taurus,and the farthest genetic relationship with Nanorana parkeri.The molecular formula of the protein encoded by the gene was Csss H1324 N234 O274 S12,the number of atoms was 2 702,the theoretical isoelectric point was 4.85,and the instability coefficient and the total average hydrophilicity were 37.22 and-0.772,respectively.which belonged to stable hydrophilic protein.MYL9 protein was a non-secreted protein without signal peptide and transmembrane helix structure.It was mainly localized in mitochondria,nucleus and cytoplasm,and had 17 specific phosphorylation sites.The secondary structure of MYL9 protein was mainly composed of alpha helix,random coil,beta turn and extended chain,and the predicted tertiary structure was consistent with the secondary structure.Real-time quantitative PCR results showed that MYL9 gene was expressed in heart,liver,spleen,lung,kidney,muscle,fat and testis,and the expression level in liver and muscle was significantly higher than that in other tissues(P＜0.05).[Conclusion]The CDS region of MYL9 gene in Meiren yak was successfully cloned,and the biological function and tissue expression characteristics of MYL9 gene were explored.The results provided a reference for further research on the functional characteristics of MYL9 gene in yak.
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