To investigate the regulation of glucose homeostasis in Micropterus salmoides, we cloned genes gp1, gp2, pepck1, pepck2, hk, pfk, and g6p encoding 878, 842, 635, 624, 918, 780, and 338 amino acid residues. We also examined the phylogenetic tree, tissue distribution, and effect of fasting on the expression of these genes. Results from multiple protein sequence alignments and phylogenetic analysis showed that these genes shared high levels of identity with their orthologs in other vertebrates. Tissue profiles of these genes revealed enrichment in the liver and muscle, but with unique tissue expression differences. For example, gp1 was highly expressed in the muscle, followed by the liver, and the lowest expression level was observed in the spleen. gp2 was also highly expressed in the muscle and abundant in the heart and liver. pepeck1 was mainly expressed in the liver and intestines, with the lowest mRNA levels in the brain, spleen, kidney, and adipose tissue. pepck2 was more significantly expressed in the liver and brain than in the other tissues, whereas the lowest mRNA levels were found in the spleen and gills. Additionally, hk and g6p were most abundantly expressed in the liver, and pfk in the liver and heart. The g6p gene was also the most abundantly expressed in the liver. The muscle, intestines, and adipose tissue were also enriched with an abundance of g6p. The highest expression levels of pfk were found in the liver and heart, followed by the intestines and gills. We examined the effect of fasting on the mRNA levels of these genes in Micropterus salmoides. The results showed that hepatic gp2, pepck2, and g6p mRNA levels were gradually up-regulated with increasing fasting time, peaking at 24 h after fasting. However, gp1, hk, and pfk exhibited diverse gene expression patterns, with transcription first increasing and then decreasing with the extension of fasting time, peaking at 6 h. Hepatic pepck1 was not significantly affected throughout the fasting period. In the muscle, gp1, gp2, and g6p mRNA expression increased significantly at 12 h, with pepck1 showing the lowest expression at 0 h and the highest expression at 12 and 24 h. After fasting for 24 h, pepck2 showed the highest mRNA level. Muscular hk and pfk showed a similar expression pattern with hepatic gp1, hk, and pfk, increasing and then decreasing with fasting time. Overall, these findings provide groundwork for future studies focusing on glucose homeostasis regulation in Micropterus salmoides.