Molecular cloning of DNA complementary to mRNA of rat liver serine dehydratase.
A cDNA clone containing sequences complementary to the mRNA cording for rat hepatic serine dehydratase was isolated to study the multihormonal regulation of this enzyme. Serine dehydratase mRNA was partially purified (50-fold enrichment, 8.2% of the total mRNA activity) from the liver of rats fed high protein diet by polysome immunoadsorption followed by oligo(dT)-cellulose column chromatography. This preparation was used as template for synthesis of cDNA. Double-stranded cDNA sequences were inserted into the plasmid pBR322 and cloned in Escherichia coli DH1. Of 860 transformants screened, 6 clones containing DNA complementary to serine dehydratase mRNA were identified by differential colony hybridization and hybrid-selected translation. The length of serine dehydratase mRNA was estimated to be 1,500 bases by Northern blot analysis. One cloned cDNA comprised about 1,000 base pairs, or 65% of the length of the mRNA. The amount of the mRNA was greatly increased in the liver of rats given high protein diet.
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D004044:Dietary Proteins / Q000008:administration & dosage / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli; D012695:L-Serine Dehydratase / Q000235:genetics*; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011132:Polyribosomes / Q000032:analysis; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats
D004044:Dietary Proteins; D012333:RNA, Messenger; D004247:DNA; D012695:L-Serine Dehydratase