The stimulatory effect of cyclic adenosine 3'5'-monophosphate on DNA-directed synthesis of beta-galactosidase in a cell-free system.
A cell-free system allowing for synthesis of beta-galactosidase enzymatic activity has been developed. This system requires DNA containing the beta-galactosidase gene, a cell-free extract of Escherichia coli bacteria, and the low-molecular-weight components necessary for transcription of the DNA and translation of the resulting messenger RNA. Such a system is useful for studying enzyme synthesis, as well as its regulation. The gene for beta-galactosidase is part of the lac operon whose expression is under the control of the lac repressor. In whole cells the lac repressor inhibits almost all of the gene expression for beta-galactosidase. In the cell-free system, we had previously been able to repress about half the gene expression. Adding cyclic adenosine 3'5'-monophosphate to the cell-free system improved the yield of beta-galactosidase enzymatic activity by 8 to 30 times and the efficiency of repression from 50 to 95 per cent.
D000227:Adenine Nucleotides / Q000494:pharmacology*; D002247:Carbon Isotopes; D002474:Cell-Free System; D000242:Cyclic AMP / Q000494:pharmacology; D004269:DNA, Bacterial; D004926:Escherichia coli; D005696:Galactosidases / Q000096:biosynthesis*; D007930:Leucine / Q000378:metabolism; D008722:Methods; D009876:Operon; D013489:Suppression, Genetic
D000227:Adenine Nucleotides; D002247:Carbon Isotopes; D004269:DNA, Bacterial; D000242:Cyclic AMP; D005696:Galactosidases; D007930:Leucine