Increased expression of a eukaryotic gene in Escherichia coli through stabilization of its messenger RNA.
The expression of a cloned eukaryotic gene [catabolic dehydroquinase (3-dehydroquinate hydro-lyase, EC 4.2.1.10) (qa-2+) from Neurospora crassa] is dramatically increased (as much as 100-fold) in Escherichia coli strains deficient in polynucleotide phosphorylase (pnp) (polynucleotide: orthophosphate nucleotidyltransferase, EC 2.7.7.8) and RNase I (rna). The increased expression is controlled primarily by the absence of polynucleotide phosphorylase and appears to be specific for the eukaryotic gene. No increase in the specific activity of either chromosomal or plasmid-borne prokaryotic genes has been observed. In polynucleotide phosphorylase-deficient strains of E. coli the half-life of plasmid (pVK88, ampr qa-2+)-encoded mRNAs increases from 1.0 to 2.8 min. This increase must be due primarily to stabilization of the aq-2 mRNA because no increase in the half-lives of pBR322 vehicle mRNAs was observed in polynucleotide phosphorylase-deficient strains. These results suggest that there are inherent structural differences between prokaryotic and eukaryotic mRNAs.
D004274:DNA, Recombinant; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D006836:Hydro-Lyases / Q000235:genetics*; D009492:Neurospora crassa / Q000201:enzymology / Q000235:genetics; D010957:Plasmids; D011117:Polyribonucleotide Nucleotidyltransferase / Q000378:metabolism; D011801:Quinic Acid / Q000031:analogs & derivatives; D012333:RNA, Messenger / Q000378:metabolism*; D012260:Ribonucleases / Q000378:metabolism
D004274:DNA, Recombinant; D012333:RNA, Messenger; D011801:Quinic Acid; D011117:Polyribonucleotide Nucleotidyltransferase; D012260:Ribonucleases; D006836:Hydro-Lyases; C020000:3-dehydroquinate dehydratase