Expression of the gene for ornithine decarboxylase of Saccharomyces cerevisiae in Escherichia coli

1985 ◽  
Vol 5 (1) ◽  
pp. 161-166
Author(s):  
W A Fonzi ◽  
P S Sypherd
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Ornithine Decarboxylase ◽  
Blot Analysis ◽  
Single Copy ◽  
Decarboxylase Activity ◽  
E Coli ◽  
Ornithine Decarboxylase Activity ◽  
Reduced Frequency ◽  
Diploid Cells

Diploid cells of Saccharomyces cerevisiae homozygous for the spe1A mutation, which eliminates ornithine decarboxylase activity, were found to sporulate at a greatly reduced frequency in the absence of polyamines. Plasmids which complement the spe1A mutation were isolated by their ability to restore sporulation competence to these cells. Three distinct plasmids were isolated. Each plasmid insert overlapped the same 8.0-kilobase region, and each plasmid restored ornithine decarboxylase activity to spe1A mutants. These plasmids also conferred ornithine decarboxylase activity to Escherichia coli EWH319 from which the ornithine decarboxylase gene is deleted. The plasmid-encoded activity expressed in E. coli resembled S. cerevisiae ornithine decarboxylase in its kinetic characteristics, indicating that the yeast ornithine decarboxylase gene was cloned. Southern blot analysis suggested that ornithine decarboxylase is a single-copy gene in S. cerevisiae. A single 2.1-kilobase transcript was demonstrated by Northern blot analysis.

scholarly journals Expression of the gene for ornithine decarboxylase of Saccharomyces cerevisiae in Escherichia coli.

1985 ◽  
Vol 5 (1) ◽  
pp. 161-166 ◽  
Author(s):  
W A Fonzi ◽  
P S Sypherd
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Ornithine Decarboxylase ◽  
Blot Analysis ◽  
Single Copy ◽  
Decarboxylase Activity ◽  
E Coli ◽  
Ornithine Decarboxylase Activity ◽  
Reduced Frequency ◽  
Diploid Cells

Diploid cells of Saccharomyces cerevisiae homozygous for the spe1A mutation, which eliminates ornithine decarboxylase activity, were found to sporulate at a greatly reduced frequency in the absence of polyamines. Plasmids which complement the spe1A mutation were isolated by their ability to restore sporulation competence to these cells. Three distinct plasmids were isolated. Each plasmid insert overlapped the same 8.0-kilobase region, and each plasmid restored ornithine decarboxylase activity to spe1A mutants. These plasmids also conferred ornithine decarboxylase activity to Escherichia coli EWH319 from which the ornithine decarboxylase gene is deleted. The plasmid-encoded activity expressed in E. coli resembled S. cerevisiae ornithine decarboxylase in its kinetic characteristics, indicating that the yeast ornithine decarboxylase gene was cloned. Southern blot analysis suggested that ornithine decarboxylase is a single-copy gene in S. cerevisiae. A single 2.1-kilobase transcript was demonstrated by Northern blot analysis.

scholarly journals Difluoromethylornithine irreversibly inactivates ornithine decarboxylase of Pseudomonas aeruginosa, but does not inhibit the enzymes of Escherichia coli

1981 ◽  
Vol 200 (1) ◽  
pp. 69-75 ◽  
Author(s):  
A Kallio ◽  
P P McCann
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Ornithine Decarboxylase ◽  
Arginine Decarboxylase ◽  
Decarboxylase Activity ◽  
Dependent Manner ◽  
Irreversible Inhibitor ◽  
E Coli ◽  
Ornithine Decarboxylase Activity

DL-alpha-Difluoromethylornithine, an enzyme-activated irreversible inhibitor of eukaryotic ornithine decarboxylase and consequently of putrescine biosynthesis, inhibited ornithine decarboxylase in enzyme extracts from Pseudomonas aeruginosa in a time-dependent manner t1/2 1 min, and also effectively blocked the enzyme activity in situ in the cell. Difluoromethylornithine, however, had no effect on the activity of ornithine decarboxylase assayed in enzyme extracts from either Escherichia coli or Klebsiella pneumoniae. However, the presence of the inhibitor in cell cultures did partially lower ornithine decarboxylase activity intracellularly in E. coli. Any decrease in the intracellular ornithine decarboxylase activity observed in E. coli and Pseudomonas was accompanied by a concomitant increase in arginine decarboxylase activity, arguing for a co-ordinated control of putrescine biosynthesis in these cells.

Biosynthetic ornithine and arginine decarboxylases: correlation of rates of synthesis with activities in Escherichia coli during exponential growth and following nutritional shift-up

1982 ◽  
Vol 28 (8) ◽  
pp. 945-950
Author(s):  
Stephen M. Boyle ◽  
Kazuo Adachi
Keyword(s):  
Escherichia Coli ◽  
Steady State ◽  
Ornithine Decarboxylase ◽  
Biological Activities ◽  
Decarboxylase Activity ◽  
Biosynthetic Enzyme ◽  
Guanosine Tetraphosphate ◽  
E Coli ◽  
K 12

Whether guanosine tetraphosphate (ppGpp) has a role in the regulation of the putrescine biosynthetic enzyme, ornithine decarboxylase, in Escherichia coli is controversial. Different laboratories have reported either direct or indirect correlations between ppGpp levels and ornithine decarboxylase activity using different in vivo conditions. In this report, using conditions in vivo to modulate ppGpp levels, experiments are described which bear on the controversy. The rates of synthesis and biological activities of the biosynthetic ornithine and arginine decarboxylases (ODC and ADC) were measured in E. coli K-12 during experimental growth and during nutritional shift-up. There were good correlations between changes in their respective activities and the rates of synthesis of these enzymes during steady state or shift-up. ODC activity or rate of synthesis changed directly in concert with ppGpp levels, while ADC activity or rate of synthesis changed inversely with ppGpp levels. These observations support the contention that ppGpp does not inhibit ODC activity.

Ornithine Decarboxylase Activity in Cultured Endothelial Cells Stimulated by Serum, Thrombin and Serotonin

1978 ◽  
Vol 39 (02) ◽  
pp. 496-503 ◽  
Author(s):  
P A D’Amore ◽  
H B Hechtman ◽  
D Shepro
Keyword(s):  
Endothelial Cells ◽  
Ornithine Decarboxylase ◽  
Decarboxylase Activity ◽  
Aortic Endothelial Cells ◽  
Ornithine Decarboxylase Activity ◽  
Rate Limiting Step ◽  
Bovine Aortic Endothelial Cells ◽  
Limiting Step ◽  
Rate Limiting ◽  
Serum Serotonin

SummaryOrnithine decarboxylase (ODC) activity, the rate-limiting step in the synthesis of polyamines, can be demonstrated in cultured, bovine, aortic endothelial cells (EC). Serum, serotonin and thrombin produce a rise in ODC activity. The serotonin-induced ODC activity is significantly blocked by imipramine (10-5 M) or Lilly 11 0140 (10-6M). Preincubation of EC with these blockers together almost completely depresses the 5-HT-stimulated ODC activity. These observations suggest a manner by which platelets may maintain EC structural and metabolic soundness.

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