Abstract: The aim was to clone the CDS region sequence of Yanbian cattle UCP1 (Uncoupling protein-1), use RT-PCR technology and gene cloning to obtain Yanbian cattle UCP1 gene, conduct homology comparison with other species and construct a phylogenetic tree, using bioinformatics methods to predict the physical and chemical properties of UCP1 encoded protein, potential phosphorylation sites and other properties, and use Quantitative Real-time PCR (qPCR) method to detect the expression abundance of UCP1 gene in different tissues of Yanbian cattle. The results showed that the CDS region of Yanbian cattle UCP1 gene was 749 bp in length, encoding 249 amino acids. The homology comparison revealed that the UCP1 gene of Yanbian cattle had 99.47% homology with Indian cattle. The phylogenetic tree showed that Yanbian cattle and Indian cattle had the closest genetic relationship and the farthest related to South American alpaca. The UCP1 protein lacked stability, the fat solubility coefficient was 91.53, the half-life of reticulocytes in vitro was 30 h, and the total average hydrophilicity was equal to 0.212, which was predicted to have a certain degree of hydrophilicity. The secondary structure was a mixed protein composed of α-helix (54.03%), extended chain (12.90%), random coil (27.42%) and β-turn (5.65%). The analysis of phosphorylation sites and glycosylation sites showed that UCP1 had a total of 21 phosphate sites, 4 potential O-glycosylation sites, and 3 N-glycosylation potential sites. The encoded product of UCP1 gene had 0 transmembrane helix (TMHs) structure, the predicted value of the amino acid residues of the transmembrane helix was 19.008 57, and the predicted value of the transmembrane helix of the first 60 amino acids of the protein was 10.980 09, which was the total probability of being located on the cytoplasmic side of the membrane 22.995% and was a non-secreted protein. Real-time fluorescent quantitative PCR (RT-PCR) showed that the expression level of UCP1 gene in Yanbian beef fat and small intestine was relatively high, while the expression level in muscle and heart was low. The experimental results could provide a reference for further research on the function of this gene.

Key words: Yanbian cattle, UCP1 gene, Cloning, Bioinformatics, Tissue mRNA expression

CLC Number: 

• Q78

• S826