Effects of temperature and novobiocin on the expression of calf prochymosin gene and on plasmid copy number in recombinant Escherichia coli
Language English Country United States Media print
Document type Journal Article
PubMed
9569630
DOI
10.1007/bf02815545
Knihovny.cz E-resources
- MeSH
- Chymosin biosynthesis genetics MeSH
- Escherichia coli genetics MeSH
- Gene Expression MeSH
- Gene Dosage MeSH
- Topoisomerase II Inhibitors MeSH
- Novobiocin pharmacology MeSH
- Plasmids drug effects genetics MeSH
- Enzyme Precursors biosynthesis genetics MeSH
- Recombinant Proteins biosynthesis MeSH
- Cattle MeSH
- DNA, Superhelical drug effects MeSH
- Hot Temperature * MeSH
- Animals MeSH
- Check Tag
- Cattle MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Chymosin MeSH
- Topoisomerase II Inhibitors MeSH
- Novobiocin MeSH
- Enzyme Precursors MeSH
- prorennin MeSH Browser
- Recombinant Proteins MeSH
- DNA, Superhelical MeSH
Escherichia coli strain HB101 harboring an expression plasmid bearing calf prochymosin gene under the control of the tac promoter was grown in the presence of IPTG with or without novobiocin at 28 and 40 degrees C, respectively. The differential rates of synthesis of prochymosin inclusions, and, for comparison, of beta-lactamase and beta-galactosidase, as well as plasmid copy number, were determined during the first hours of steady state growth. At 28 degrees C the induced expression of prochymosin gene was almost blocked. Addition of novobiocin did not alleviate this effect. In fact, it strengthened it, and we conclude that both these additive inhibitory effects are a consequence of the decrease in negative superhelical tension of plasmid DNA to an insufficient level. At 40 degrees C the differential rate of prochymosin synthesis was markedly enhanced. Since the copy number of the expression plasmid increased approximately to the same extent, we conclude that an increase in gene dose is the cause. The stimulation of cloned heterologous gene expression at 40 degrees C and inhibition at 28 degrees C may be conveniently used in biotechnological-scale cultivations of some recombinant bacteria.
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