Abstract: Objective: In this paper, we obtained the NOT5 gene from Saccharomyces uvarum by transcriptome analysis. Analysis of its biological information can lay the foundation for the later study of the role of the gene. Methods: The primary structure, secondary structure, tertiary structure, and structural domains of the encoded protein were predicted using online analysis tools such as COILS Server, SOPMA, and Alpha Fold. Results: The length of the NOT5 gene was 1446 bp, encoding 481 amino acids, and it was located on chromosome XVI 690107~691789. The genetic relationship analysis showed that this gene was the closest to Saccharomyces eubayanus NOT5 like protein XP018219088.1, and had high homology. Its molecular formula was C_2493H3848N₆₅₄O₇₉₅S₁₉, and its molecular mass was 56311.02. It was an unstable hydrophilic protein. The protein encoded by this gene did not contain a signal peptide, had no transmembrane region, had a coiled helix region, and was subcellularly localized on the mitochondria in the cytoplasm. The secondary structure of the protein was dominated by random coils. Pfam Not3 and Pfam NOT2_3_5 domains were predicted. Yeast Not1-Not2-Not5 (4by6.1.C) was used as the template to construct the tertiary structure of NOT5 protein, and the sequence consistency between the two could reach 89.88%. Conclusion: The structure of the protein encoded by the NOT5 gene was unstable, and would interconnect translation and transcription in the cell.