Cloning and expression analysis of hepatic lipase from Pampus argenteus

摘要

Hepatic lipase (HL) is a key enzyme in endogenous triglyceride metabolism in blood circulation. The molecular mechanism and function of Pampus argenteus HL gene was investigated by cloning the full-length cDNA of HL using PCR and RACE. The temporal and spatial expression characteristics of HL in different tissues and growth stages of P. argenteus and its localization in the liver were examined using real-time PCR and in situ hybridization, and liver crude fat content was determined. The full-length cDNA of P. argenteus HL gene was 1836 bp (GenBank No. KU497716); the 1485 bp ORF encodes 494 amino acids. P. argenteus HL protein had the highest homology with Siniperca chuatsi HL protein. In P. argenteus, HL was highly expressed in the liver and scarcely expressed in other tissues. HL expression in P. argenteus initially increased and then decreased at different growth stages (1-5, 5-15, 15-30, 30-50, 50-100 and >100 g), with the highest expression at 30-50 g fish weight, which was positively correlated with liver fat content. In situ hybridization showed that, in liver cells, HL was highly and weakly expressed in the cytoplasm and nucleolus, respectively, and not expressed in the nuclear interstitium.