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Molecular Cloning of Rat Genomic DNA for Epinephrine Synthesizing Enzyme, Phenylethanolamine N-Methyltransferase

Molecules and Cells 1990년 1권 1호 p.37 ~ 41
 ( Suh Yoo-Hun ) - Seoul National University College of Medicine Department of Pharmacology

 ( Kim Hun-Sik ) - Seoul National University College of Medicine Department of Pharmacology
 ( Kim Sung-Soo ) - Seoul National University College of Medicine Department of Pharmacology
 ( Choi Woong ) - Seoul National University College of Medicine Department of Pharmacology
 ( Park Chan-Woong ) - Seoul National University College of Medicine Department of Pharmacology

Abstract


To obtain information on the structure and expression regulation of the gene encoding epinephrine synthesizing enzyme, phenylethanolarrune N-methyltransferase (PNMT), which plays a vital role in expression of adrenergic phenotypes, bovine PNMT cDNA was used as a probe to screen a rat genomic library constructed in EMBL3 replacement vector at BamHI sites. About 106 phage plaques were examined and one clone showing strong hybridization signals on 3 repetitive screening procedures was isolated. The recombinant DNA purified from this rat genomic clone hybridized to both 5'- and 3'-end specific bovine PNMT cDNA probes. Complete BamHI digestion of this recombinant DNA yields 3 insert DNA fragments of 6.7 kb, 5.0 kb and 1.5 kb. Southern hybridization analysis showed that the 6.7-kb insert fragment contains the sequence homologous to bovine PNMT cDNA, and it was mapped with some restriction enzymes. The unique KpnI site located at approximately 2.5 kb from the 5' end of this insert fragment separates the exons of the rat PNMT gene cloned in this work.

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