Cloning of DNA complementary to ornithine aminotransferase mRNA.
Free polysomes were isolated from the livers of rats maintained on a 60% casein diet to induce ornithine aminotransferase mRNA. Ornithine aminotransferase-synthesizing polysomes were immunoadsorbed using monospecific, affinity-purified antibody and Staphylococcus aureus cells. Poly(A+)RNA prepared from these polysomes by oligo(dT)-cellulose chromatography was used as a template for the synthesis of double-stranded cDNA by reverse transcriptase. Using the deoxyguanosine-5'-triphosphate-deoxycytidine-5'-triphosphate tailing method and pBR322 as a vector, recombinant molecules were produced and used to transform Escherichia coli. Two clones containing DNA complementary to ornithine aminotransferase mRNA were identified by colony hybridization and hybrid-arrest translation. A partial restriction endonuclease map of the ornithine aminotransferase cDNA inserts was constructed which spanned 1066 base pairs.
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004262:DNA Restriction Enzymes; D008099:Liver / Q000201:enzymology*; D008297:Male; D009953:Ornithine-Oxo-Acid Transaminase / Q000235:genetics*; D011061:Poly A / Q000235:genetics; D011132:Polyribosomes / Q000201:enzymology; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D011919:Rats, Inbred Strains; D013211:Staphylococcus aureus / Q000235:genetics; D000637:Transaminases / Q000235:genetics*
D012333:RNA, Messenger; D011061:Poly A; D004247:DNA; D000637:Transaminases; D009953:Ornithine-Oxo-Acid Transaminase; D004262:DNA Restriction Enzymes