Bioinformatics and Expression Characteristics Analysis of MGP Gene in Qinchuan Cattle
[Objective]This study was aimed to predict the fundamental functions of matrix Gla protein(MGP),reveal the tissue expression patterns of MGP gene in Qinchuan cattle and its expression characteristics during different stages of preadipocyte differentiation,so as to provide a theoretical foundation for studying the regulatory mechanisms of MGP gene in the adipose development of Qinchuan cattle.[Method]The similarity alignment and phylogenetic tree construction of MGP protein in various species were conducted,and the physicochemical properties and subcellular localization of bovine MGP protein were predicted using bioinformatics online software.The differentiation effect of preadipocytes in Qinchuan cattle was examined using oil Red O staining.The expression of MGP gene in different tissues of Qinchuan cattle and its characteristics during different stages of preadipocyte differentiation were detected by Real-time quantitative PCR.[Result]The similarity alignment results showed that the similarity of MGP protein was the highest(99％)between Bos taurus and Capra hircus,while the similarity was the lowest(31％)with Danio rerio.The phylogenetic tree revealed that Bos taurus and Capra hircus shared the closest relationship,whereas the relationship was the most distant with Danio rerio.MGP gene encoded 103 amino acids with a theoretical isoelectric point of 9.27,indicating alkalinity.MGP was considered a hydrophilic protein with a total average hydrophobicity index of-0.630.MGP was classified as a secretory protein due to the presence of a Sec/SP Ⅰ-type signal peptide and the absence of transmembrane structures.Additionally,MGP protein included 5 serine and 4 threonine phosphorylation sites,and it possessed a GLA_domain containing γ-carboxyglutamic acid(Gla).The alpha helix constituted the highest percentage of the secondary structure at 55.34％.The oil Red O staining confirmed the well-differentiated state of isolated and cultured preadipocytes.Real-time quantitative PCR results showed that the expression of MGP gene in heart of Qinchuan cattle was significantly higher than that in other tissues(P＜0.05).Elevated expression were also observed in large intestine and perirenal fat,while the expression in small intestine was significantly lower than that in other tissues(P＜0.05).MGP gene exhibited a trend of initial upregulation followed by downregulation during different stages of preadipocyte differentiation,with significantly higher expression on the 2nd day of preadipocyte induction compared with other days(P＜0.05),and significantly lower expression on the 10th day compared with other days(P＜0.05).[Conclusion]This study predicted the basic structure and function of MGP protein.The expression of MGP gene was the highest in heart of Qinchuan cattle and was the lowest in small intestine.MGP gene exhibited a trend of initial upregulation followed by downregulation during different stages of preadipocyte differentiation.
万方数据
维普
