关键词: CYP2J5基因, 超表达载体, 生物信息学

Abstract: This study firstly analyzed the tissue differential expression of CYP2J5 gene by qRT-PCR, and then used bioinformatics tools to predict and analyze the structure and function of mouse CYP2J5. To further explore and verify the biological function and its effects on lipid metabolism and sex hormone synthesis, the overexpression vector of CYP2J5 gene was constructed based on genetic engineering technology, and the constructed vector was confirmed by Western blot which can express CYP2J5 gene correctly. Finally, the constructed CYP2J5 overexpression vector was introduced into TM3 cells, and the expression of lipid metabolism and sex hormone synthesis related genes were analyzed by qRT-PCR. The results showed that the gene was highly expressed in the liver and kidney, and it was a soluble cytoplasmic protein. The secondary and tertiary structure of the protein was mainly composed of α-helix and random coil, the signal peptide existed and there was a transmembrane region between the 13th to 35th and 75th to 97th amino acids, whose main functions were involved in biological regulation processes such as cell lipid metabolism and fatty acid metabolism. After overexpression of the gene in TM3 cells, the key genes in the lipid metabolism and sex hormone pathways were significantly altered. The results of this study provid basic data on bioinformatics and molecular biology for the research and development gene function of CYP2J5.

Key words: CYP2J5 gene, overexpression vector, bioinformatic analysis