Molecular cloning and amplification of the adenylate cyclase gene.
A segment of DNA containing cya, the gene for adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1], has been isolated from Salmonella typhimurium. The phage lambda gt4 was used as a cloning vector and adenylate cyclase-positive hybrid phages were isolated that complemented adenylate cyclase-negative bacteria. The cloned DNA fragment encodes a polypeptide of molecular weight 81,000 that gives rise to adenylate cyclase activity. This protein represents a functional mutant of the bacterial adenylate cyclase. When the cya gene was amplified by inserting into a multicopy plasmid, the enzyme activity was overproduced 20-fold, but the cyclic AMP level increased only 60%, suggesting several probable regulatory mechanisms. Overproduction of enzymes by recombinant DNA techniques can be a useful probe of relationships in the metabolizing organism in vivo.
D000262:Adenylyl Cyclases / Q000235:genetics*; D010582:Bacteriophage lambda / Q000235:genetics; D002874:Chromosome Mapping; D003001:Cloning, Molecular / Q000379:methods; D000242:Cyclic AMP / Q000378:metabolism; D005784:Gene Amplification; D005786:Gene Expression Regulation; D005796:Genes; D008970:Molecular Weight; D009876:Operon; D010957:Plasmids; D012486:Salmonella typhimurium / Q000235:genetics*
D000242:Cyclic AMP; D000262:Adenylyl Cyclases