The cloning and expression of a murine triacylglycerol hydrolase cDNA and the structure of its corresponding gene

A novel murine cDNA for triacylglycerol hydrolase (TGH), an enzyme that is involved in mobilization of triacylglycerol from storage pools in hepatocyt es, has been cloned and expressed. The cDNA consists of 1962 bp with an ope n reading frame of 1695 bp that encodes a protein of 565 amino acids. Murin e TGH is a member of the CESIA class of carboxylesterases and shows a signi ficant degree of identity to other carboxylesterases from rat, monkey and h uman. Expression of the cDNA in McArdle RH7777 hepatoma cells showed a 3-fo ld increase in the hydrolysis of p-nitrophenyl laurate compared to vector-t ransfected cells. The highest expression of TGH was observed in the livers of mice, with lower expression in kidney, heart, adipose and intestinal (du odenum/jejunum) tissues. The murine gene that encodes TGH was cloned and ex on-intron boundaries were determined. The gene spans approx. 35 kb and cont ains 14 exons. The results will permit future studies on the function of th is gene via gene-targeting experiments and analysis of transcriptional regu lation of the TGH gene. (C) 2001 Published by Elsevier Science B.V.