Construction of Staphylococcus plasmid vector pCA43 conferring resistance to chloramphenicol, arsenate, arsenite and antimony

Kreutz, Birgit; Götz, Friedrich

doi:10.1016/0378-1119(84)90226-9

Cited by 37 publications

(22 citation statements)

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“…pCA43 is a recombinant plasmid consisting of the vector pC194 and a DNA fragment containing the arsenical resistance genes from pSX267 (21). pCA44 is a deletion product of pCA43 lacking the pSX267-derived 1.2-kb PvuII-ScaI fragment (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…S. carnosus plasmid pC194 (17) is a Staphylococcus vector plasmid. pCA43 is a pC194 replicon containing the arsenical resistance genes of S. xylosus DSM 20267 (21). pLipPl is a pC194 replicon containing the S. hyicus lipase gene (lip) (12).…”

Section: Methodsmentioning

confidence: 99%

“…The resulting plasmid, pCA43 (21), confers resistance to chloramphenicol, arsenate, arsenite, and antimonite. Insertional inactivation studies showed that the arsenical resistance * Corresponding author.…”

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confidence: 99%

“…determinants are composed of at least two genes, one of which encodes both the arsenite and antimonite resistances. Arsenate resistance occurs only when both genes are intact (21).…”

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confidence: 99%

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Expression and regulation of the antimonite, arsenite, and arsenate resistance operon of Staphylococcus xylosus plasmid pSX267

et al. 1992

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The arsenate, arsenite, and antimonite resistance region of the Staphylococcus xylosus plasmid pSX267 was subcloned in Staphylococcus carnosus. The sequenced DNA region revealed three consecutive open reading frames, named arsR, arsB, and arsC. Expression studies in Escherichia coli with the bacteriophage T7 RNA polymerase-promoter system yielded three polypeptides with apparent molecular weights of 8,000, 35,000, and 15,000, which very likely correspond to ArsR, ArsB, and ArsC, respectively. ArsB was distinguished by its overall hydrophobic character, suggesting a membrane association. The arsenate, arsenite, and antimonite resistance was shown to be inducible by all three heavy metal ions. Inactivation of the first gene, arsR, resulted in constitutive expression of resistance. Similar results were obtained with transcriptional fusions of various portions of the ars genes with a lipase reporter gene, indicating a function of ArsR as a negative regulator of a putative promoter in front ofarsR. The inactivation ofarsR also resulted in reduction of resistance to arsenite and antimonite, while arsenate resistance was unaffected. The three ars genes conferred arsenite resistance in E. coli and arsenite as well as arsenate resistance in BaciUus subtilis.Plasmid-mediated resistance to arsenate, arsenite, and antimonite salts in Staphylococcus aureus was first described by Novick and Roth (29). The resistance genes are normally found on plasmids encoding penicillinase and various heavy metal resistance determinants, such as resistance to Cd2' and Hg'. The arsenical resistance genes on plasmid p1258 are composed of at least two loci (23), which are clustered (28). Studies on the arsenical resistance operon of the Escherichia coli conjugative R-factor plasmid R773 and the S. aureus plasmid pI258 revealed that the resistance determinants from both organisms are inducible by arsenate, arsenite, or antimonite. The mechanism of plasmid-mediated arsenate resistance in both E. coli and S. aureus is based on an efflux system (25,42), which, at least in E. coli, functions as an ATPase. It was demonstrated that, at least in E. coli, arsenite is translocated similarly to arsenate (32).Arsenate and arsenite resistances are common in other staphylococcal species. An investigation of a collection of Staphylococcus xylosus strains isolated from soybean oil meal showed that more than 90% were arsenate and arsenite resistant and that the resistance genes were plasmid encoded. One strain, S. xylosus DSM 20267, harboring a 29.5-kb plasmid, designated pSX267, was studied in more detail. In searching for a pSX267-associated resistance to different antibiotics, no indication of plasmid-bome resistance to any of 63 antibacterial drugs tested was detected in this strain. Heteroduplex analysis between pSX267 and p1258 revealed only high DNA homology within the arsenical resistance genes and the replication region (14).The arsenical resistance genes of pSX267 have been subcloned in Staphylococcus carnosus by using pC194. The resulting plasmid, p...

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

“…determinants are composed of at least two genes, one of which encodes both the arsenite and antimonite resistances. Arsenate resistance occurs only when both genes are intact (21).…”

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confidence: 99%

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Expression and regulation of the antimonite, arsenite, and arsenate resistance operon of Staphylococcus xylosus plasmid pSX267

et al. 1992

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“…S. carnosus and S. aureus were grown in tryptic soy broth medium (Invitrogen) or LB medium (1% tryptone, 0.5% yeast extract, and 0.5% NaCl). The plasmid vectors pCA44 (7,8) and the xylose-inducible pTX15 (9, 10) were used; for induction in the latter case, xylose was added to the medium to a final concentration of 0.5%. PCR was carried out with Vent polymerase (New England Biolabs) or Expand TM Long Template PCR System (Roche Applied Science).…”

Section: Methodsmentioning

confidence: 99%

Structure and Biosynthesis of Staphyloxanthin from Staphylococcus aureus

Pelz

Wieland

Putzbach

et al. 2005

Journal of Biological Chemistry

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307

280

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Protein Production inStaphylococcus carnosusand other Gram‐Positive Bacteria

Freudl

2008

Protein Science Encyclopedia

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Originally published in: Production of Recombinant Proteins. Edited by Gerd Gellissen. Copyright © 2005 Wiley‐VCH Verlag GmbH & Co. KGaA Weinheim. Print ISBN: 3‐527‐31036‐4 The sections in this article are Introduction Major Protein Export Routes in Gram‐positive Bacteria The General Secretion (Sec) Pathway The Twin‐Arginine Translocation (Tat) Pathway Secretion Signals Extracytosolic Protein Folding The Cell Wall as a Barrier for the Secretion of Heterologous Proteins Degradation of Exported Proteins by Cell‐associated and Secreted Proteases Staphylococcus carnosus General Description Microbiological and Molecular Biological Tools S. carnosus as Host Organism for the Analysis of Staphylococcal‐related Pathogenicity Aspects Secretory Production of Heterologous Proteins by S. carnosus The Staphylococcus hyicus Lipase: Secretory Signals and Heterologous Expression in S. carnosus Use of the Pre‐pro‐part of the S. hyicus Lipase for the Secretion of Heterologous Proteins in S. carnosus Process Development for the Secretory Production of a Human Calcitonin Precursor Fusion Protein by S. carnosus Surface Display on S. carnosus Appendix

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Construction of Staphylococcus plasmid vector pCA43 conferring resistance to chloramphenicol, arsenate, arsenite and antimony

Cited by 37 publications

References 9 publications

Expression and regulation of the antimonite, arsenite, and arsenate resistance operon of Staphylococcus xylosus plasmid pSX267

Expression and regulation of the antimonite, arsenite, and arsenate resistance operon of Staphylococcus xylosus plasmid pSX267

Structure and Biosynthesis of Staphyloxanthin from Staphylococcus aureus

Protein Production inStaphylococcus carnosusand other Gram‐Positive Bacteria

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