ELSEVIER Biochimica et Biophysica Acta 1309 (1996) 21-24 BIOCHIMICA ET BIOPHYSICA ACTA BBI Short sequence-paper Cloning and expression of the cDNA for mouse NAD +-dependent 15-hydroxyprostaglandin dehydrogenase Muneaki Matsuo, Charles Mark Ensor, Hsin-Hsiung Tai * Division of Medicinal Chemisto, and Pharmaceutics, College of Pharmacy, University of Kentucky, Lexington, KY 40536, USA Received 12 January 1996; revised 5 June 1996; accepted 7 June 1996 Abstract The cDNA for mouse NAD + dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) was isolated from a lung cDNA library. The cDNA contains a 798 bp open reading frame that codes for a protein of 266 amino acids (M r 28775) which shares 87% identity with the human 15-PGDH protein. The regions that are believed to form the NAD + binding site and the active site are conserved in the mouse and human enzymes. The authenticity of the mouse cDNA was confirmed by expression of an active 15-PGDH in Escherichia coli. Northern blot analysis demonstrated that 15-PGDH mRNA is expressed primarily in lung, intestine, stomach and liver. Keywords: NAD + dependent 15-hydroxyprostaglandin dehydrogenase; cDNA; Gene expression; Lung; (Mouse) NAD+-dependent 15-hydroxyprostaglandin dehy- drogenase (15-PGDH) is considered to be a key catabolic enzyme that controls the biological activity of prostaglandins [1,2]. The 15-PGDH cDNA has been previously cloned from human placenta [3]. Although the enzyme is ubiquitous in mammalian tissues [2], the extent of structural similarity of the enzyme from different tissues and species remains unknown. In addition, little is known about the bio- synthetic regulation of 15-PGDH or its precise in vivo role. Since a 15-PGDH-deficient mouse model, created by the gene targeting technology, would be valuable for a better understanding of the in vivo role of 15-PGDH, we have cloned the cDNA for mouse lung 15-PGDH by hybridization screening * Corresponding author. Fax: + 1 (606) 2577585. The 789 bp coding sequence from the human 15-PGDH cDNA was radiolabeled and used to screen a hgtll mouse lung cDNA library (Clontech, Palo Alto, CA) by plaque hybridization [4]. One positive clone was plaque purified from 1.2 X l0 s phage and the cDNA insert was amplified by the polymerase chain reaction (PCR) using )tgtll primers which flank the insertion site of the cDNA and sequenced. Fig. 1 shows the nucleotide sequence of the mouse lung 15-PGDH cDNA and the deduced amino acid sequence. The cDNA contains the complete coding sequence plus 24 bp of 5' untranslated region and 482 bp of the 3' untranslated region. The coding sequence is 798 bp long and codes for a protein of 266 amino acids with a calculated molecular weight of 28775. In Fig. 2 the deduced amino acid se- quences of the mouse lung 15-PGDH and the human placental 15-PGDH are compared. Both contain 266 0167-4781/96/$15.00 Copyright © 1996 Elsevier Science B.V. All rights reserved. PII S0167-4781 (96)00123-6