Cloning and Expression Analysis of FTO Gene in Yak (Bos grunniens)
GU Ya-Rong1,2, MA Lan-Hua1, WANG Fu-Bin1,2, ZHANG Yong-Feng2, Yan Ping2, Pan He-Ping1,*
1 College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China; 2 Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences, Lanzhou 730050, China
Abstract：Fat mass and obesity associated gene (FTO) belongs to the non-heme plus dioxygenase superfamily and plays an important role in mammalian fat development. This study takes Qinghai Datong yak (Bos grunniens) as the research object, the CDS region of FTO gene was amplified by RT-PCR, and its function and structure were predicted by bioinformatics online softwares. At the same time, the expression of FTO gene in different stages (18 and 30 months old), different tissues (heart, liver, spleen, lung, kidney, longissimus dorsi muscle, subcutaneous adipose) and different stages of yak preadipocyte differentiation (0, 4, 8 and 12 d) were analyzed by qPCR. The results showed that the full length of FTO (GenBank No. OM640141) coding region was 1 518 bp, encoding 505 amino acids. The FTO protein mainly consisted of α-helix (44.36%), irregularly coiled (38.42%), extended chain (10.89%) and β-turned (6.34%), without transmembrane structure and signal peptide, belonging to an unstable hydrophilic protein. The results of phylogenetic tree showed that the amino acid sequence of FTO was 99.80% homologous to cattle (Bos taurus) with the closest genetic distance, and was closely related to zebu (B. indicus) and wild yak (B. mutus), and most distantly related to horse (Equus przewalskii). The results of qPCR showed that FTO expression was highest in yak heart tissue and subcutaneous adipose tissue, and lowest in spleen. The expression of FTO was significantly higher at 30 months than that at 18 months age (P<0.05). The FTO expression was found to be significantly decreased at 4, 8 and 12 d (P<0.05) during yak preadipocyte differentiation, and the FTO protein expression detected by Western blot had the same changing trend. This study can provide basic material for further research on the biological function of FTO gene in yak fat development.
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