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KMID : 0578319940040050037
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Molecules and Cells
1994 Volume.4 No. 5 p.37 ~ p.41
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Molecular Cloning of Rat Genomic DNA for Epinephrine Synthesizing Enzyme, Phenylethanolamine N-Methyltransferase
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Suh Yoo-Hun
Kim Hun-Sik
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Abstract
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To obtain information on the structure and expression regulation of the gene encoding epinephrine synthesizing enzyme, phenylethanolamine 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 10`´ 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 Y-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 Kpnl site located ar 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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