Genomic restriction endonuclease analysis and mapping of murine guanylate cyclase-A/atrial natriuretic factor receptor gene.
The membrane-bound form of guanylate cyclase represents a biologically active atrial natriuretic factor receptor (GC/ANF-R). We have constructed genomic map of murine GC-A/ANF-R gene using 17 different restriction endonucleases. The restriction mapping results indicated that murine GC-A/ANF-R gene is approximately 20 kb single copy with multiple smaller exons and bigger introns. The Kpn I and Sfu I restriction digests produced 27 kb and 35 kb fragments, respectively, which hybridized with 5'- and 3'-flanking cDNA probes. Both of these fragments should cover the entire murine GC-A/ANF-R genomic sequences. The southern blot hybridization of genomic DNA from human, rat and mouse, using murine 5'-flanking cDNA probe indicated the presence of higher variant sequences in the 5'-flanking region of GC-A/ANF-R gene among different species. The noncoding 5'-flanking probe (350 bp) hybridized only to mouse genomic DNA but not to the human or rat DNA. These sequence variations located in the noncoding 5'-flanking region of GC-A/ANF-R gene may explain the divergent evolutionary development among different species. This is the first demonstration of the restriction endonuclease digestion and genomic mapping of murine GC-A/ANF-R gene which should be valuable to the understanding of its regulation and function.
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Department of Biochemistry and Molecular Biology, Medical College, Georgia School of Medicine Augusta 30912.
Pandey
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D000818:Animals; D001011:Aorta / Q000378:metabolism; D001483:Base Sequence; D015139:Blotting, Southern; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D002999:Clone Cells; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D015342:DNA Probes; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D005091:Exons; D006162:Guanylate Cyclase / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007438:Introns; D007984:Leydig Cell Tumor / Q000378:metabolism*; D051379:Mice / Q000235:genetics*; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D051381:Rats; D017461:Receptors, Atrial Natriuretic Factor / Q000096:biosynthesis / Q000235:genetics*; D015183:Restriction Mapping; D013379:Substrate Specificity
D017931:DNA Primers; D015342:DNA Probes; D004273:DNA, Neoplasm; D004247:DNA; D004262:DNA Restriction Enzymes; D006162:Guanylate Cyclase; D017461:Receptors, Atrial Natriuretic Factor