title: Cloning of full-length preproendothelin-2 cDNA and its expression in dog.
abstract: Endothelin-2 (ET2) is a member of the endothelin family of 21-amino acid peptides with vasoconstrictive activity. We report here the molecular cloning of the canine full-length cDNA of the precursor form of ET2, prepro-ET2 (PPET2), from intestinal tissue by means of reverse transcription-polymerase chain reaction (RT-PCR) in conjunction with 5'- and 3'-rapid amplification of cDNA ends (RACE). Aside from the poly (A) tail the cDNA was found to be 1195 bp and included an open reading frame of 534 bp encoding a PPET2 polypeptide of 178 residues, in which the regions corresponding to bioactive mature ET2 peptide, an intermediate form big-ET2, and endothelin-like peptide are found. The organ distributions of PPET2 mRNA and a splicing variant were analyzed by RT-PCR. PPET2 transcript was detected in duodenum, colon, stomach, lung, liver, uterus, ovary, testis and kidney, but not in spleen. A splicing variant was found in none of the organs. Thus, based on the cloned cDNA sequence, we established a quantitative assay for dog PPET2 mRNA level using a real-time PCR system. Quantitative analysis by this method in various organs of the dog demonstrated that the dominant gene expression occurs in the intestine, with higher expression in large intestine than in small intestine.
authors.affiliation: Department of Toxicology, School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Fujimori
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Uchide
authors.forename: Kaname
authors.initials: K
authors.lastname: Saida
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Temma
authors.forename: Takushi
authors.initials: T
authors.lastname: Sasaki
authors.forename: Tai
authors.initials: T
authors.lastname: Akera
mesh_terms: D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs; D016232:Endothelins / Q000235:genetics*; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D016232:Endothelins; D011498:Protein Precursors; D012333:RNA, Messenger; C099648:preproendothelin 2