Control of tryptophan synthetase amplified by varying the numbers of composite plasmids in Escherichia coli cells

Gene
K NagahariK Sakaguchi

Abstract

Using pSC101, RSF1010, RSF2124 and RP4 plasmids as vectors and bacteriophage lambdatrpD-A60-3 DNA as a source of the Escherichia coli whole tryptophan operon, composite plasmids of pSC101-trp, RSF1010-trp, RSF2124-trp and RP4-trp were constructed in vitro with EcoRI restriction endonuclease and DNA ligase. Each composite plasmid could be maintained stably in E. coli cells. The copy number of pSC101-trp, RSF1010-trp, RSF2124-trp and RP4-trp were 4.2, 11.2, 11.9 and 1.6 per chromosome respectively. The tryptophan synthetase activities in cells containing pSC101-trp, RSF1010-trp, RSF2124-trp aand RP4-trp plasmid were found to be 2.1, 6.0, 5.0 and 2.5 times compared with the level in chromosomal trp+ cells when they were grown in a minimal medium. By partial derepression with indolylacrylic acid, the enzyme levels were elevated to 10.1, 16.3, 15.3, 12.3 times, respectively, that of the control cells. The tryptophan synthetase activities did not increase in proportion to the copy number of the plasmids, but were strongly affected by the repression system of host cells.

References

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Citations

Aug 1, 1979·Molecular & General Genetics : MGG·P CossartG N Cohen
Jun 20, 1979·Molecular & General Genetics : MGG·L J Wallace, G Wilcox
Mar 20, 1979·Molecular & General Genetics : MGG·K NagahariK Sakaguchi
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Dec 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·Y MachidaJ E Ikeda
Oct 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·M A HedigerI Zabin
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Dec 21, 2016·Microbial Cell Factories·Michael JahnSusann Müller
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Mar 1, 1979·Microbiological Reviews·B W HollowayA F Morgan

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