Cloning, Expression in Escherichia coli and Nucleotide Sequence of a Tetracycline-resistance Gene from Streptomyces rimosus

Reynes, Jean Paul; Calmels, Thierry; Drocourt, Daniel; Tiraby, Gérard

doi:10.1099/00221287-134-3-585

Cited by 28 publications

(29 citation statements)

References 37 publications

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“…Regions of complementarity of three nucleotides or more were created in each of the constructs that gave increased levels of expression, except for pIJ2242 (Fig. 2) (18,35), the synthesis of the active form of the antibiotic in the vicinity of the efflux machinery might be essential for resistance. Previous studies (3) had shown that the ORF 1 and 2 products were homologous to the that of the nodE gene of Rhizobium species; interestingly, this protein is also located on the membrane (41).…”

Section: Discussionmentioning

confidence: 99%

Overproduction and localization of components of the polyketide synthase of Streptomyces glaucescens involved in the production of the antibiotic tetracenomycin C

et al. 1991

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Three proteins, including the 13-keto acyl synthase and the acyl carrier protein, involved in the synthesis of the polyketide antibiotic tetracenomycin C by Streptomyces glaucescens GLA.0 were produced in Escherichia coli by using the T7 RNA polymerase-dependent pT7-7 expression vector. Changing the N-terminal codon usage of two of the genes greatly increased the level of protein produced without affecting mRNA levels, suggesting improvements in translational efficiency. Western immunoblot analysis of cytoplasmic and membrane fractions of S. glaucescens with antibodies raised to synthetic oligopeptides corresponding to the two presumed components of the jI-keto acyl synthase indicated that both proteins were membrane bound; one appears to be proteolytically cleaved before or during association with the membrane. The j-keto acyl synthase could be detected in stationary-phase cultures but not in rapidly growing cultures, correlating with the time of appearance of tetracenomycin C in the medium.

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Section: Discussionmentioning

confidence: 99%

Overproduction and localization of components of the polyketide synthase of Streptomyces glaucescens involved in the production of the antibiotic tetracenomycin C

et al. 1991

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“…The otrB gene encodes an integral membrane protein that is responsible for efflux of OTC from the cell (151,167,198). A third OTC resistance gene, otrC, has been cloned from S. rimosus, but the mechanism of resistance encoded by this gene has yet to be elucidated (I. S. Hunter, unpublished data).…”

Section: Cloning Of Tetracycline Resistance Genesmentioning

confidence: 99%

“…In the scientific literature, these bacteria have come to be known as the Russian (LS-T118), Zagreb (R6), and Pfizer (M4018) strains (94,117). Other strains have also been used sporadically as donors or recipients of DNA (e.g., BS-21, 907, 183, RCC, and PG3) (41,155,188,198,212), but no more data on their genetics have been published to date.…”

Section: Introductionmentioning

confidence: 99%

“…Restriction-deficient mutants (115) and broad-host-range plasmids (64,75,126,135) and phage vectors (208), including a bifunctional cosmid for use with S. rimosus, have also become available (40). These genetic tools and procedures have been applied to study the genetic instability of S. rimosus strains (49,80) and for the molecular cloning and characterization of the OTC resistance gene(s) (37,61,167,198) as well as the cloning of genes involved in OTC biosynthesis (29,36,37,150,177). These genes have also been used for the formulation of design rules for constructing hybrid aromatic polyketide synthases with a view to producing structural analogues, by combinato- rial biosynthesis, with potentially novel biological activities which are difficult to produce by traditional synthetic routes (125,148,175,177).…”

Section: Introductionmentioning

confidence: 99%

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Genetics ofStreptomyces rimosus, the Oxytetracycline Producer

Petković

Cullum

Hranueli

et al. 2006

Microbiol Mol Biol Rev

101

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SUMMARY From a genetic standpoint, Streptomyces rimosus is arguably the best-characterized industrial streptomycete as the producer of oxytetracycline and other tetracycline antibiotics. Although resistance to these antibiotics has reduced their clinical use in recent years, tetracyclines have an increasing role in the treatment of emerging infections and noninfective diseases. Procedures for in vivo and in vitro genetic manipulations in S. rimosus have been developed since the 1950s and applied to study the genetic instability of S. rimosus strains and for the molecular cloning and characterization of genes involved in oxytetracycline biosynthesis. Recent advances in the methodology of genome sequencing bring the realistic prospect of obtaining the genome sequence of S. rimosus in the near term.

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“…TetM has been shown to bind noncovalently to the ribosome and promote removal of tetracycline (81,82). On the other hand, OtrB was shown to be a membrane-bound protein responsible for reduced accumulation of tetracycline in the cell (79,83). The gene encoding OtrB lies with opposite polarity to the gene encoding the regulatory protein OxyTA1 in a similar fashion as the tetR/tetA pair, in which TetR negatively regulates expression of the efflux protein TetA (84).…”

Section: Regulatory and Resistance Genesmentioning

confidence: 99%

Oxytetracycline Biosynthesis

Pickens¹,

Tang

2010

Journal of Biological Chemistry

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Oxytetracycline (OTC) is a broad-spectrum antibiotic that acts by inhibiting protein synthesis in bacteria. It is an important member of the bacterial aromatic polyketide family, which is a structurally diverse class of natural products. OTC is synthesized by a type II polyketide synthase that generates the poly-␤-ketone backbone through successive decarboxylative condensation of malonyl-CoA extender units, followed by modifications by cyclases, oxygenases, transferases, and additional tailoring enzymes. Genetic and biochemical studies have illuminated most of the steps involved in the biosynthesis of OTC, which is detailed here as a representative case study in type II polyketide biosynthesis.

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Cloning, Expression in Escherichia coli and Nucleotide Sequence of a Tetracycline-resistance Gene from Streptomyces rimosus

Cited by 28 publications

References 37 publications

Overproduction and localization of components of the polyketide synthase of Streptomyces glaucescens involved in the production of the antibiotic tetracenomycin C

Overproduction and localization of components of the polyketide synthase of Streptomyces glaucescens involved in the production of the antibiotic tetracenomycin C

Genetics ofStreptomyces rimosus, the Oxytetracycline Producer

Oxytetracycline Biosynthesis

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