K. N. Timmis

Details der Publikationsliste

Zeitraum

1984 - 2007

Anzahl

208

Co-Autoren

Genomotyping of Pseudomonas putida strains using P. putida KT2440-based high-density DNA microarrays: implications for transcriptomics studies (2007)

Ballerstedt, H., Volkers, R.J.M., Mars, A.E., Hallsworth, J.E., Martins Dos Santos, V.A., Puchalka, J., ...

Pseudomonas putida KT2440 is the only fully sequenced P. putida strain. Thus, for transcriptomics and proteomics studies with other P. putida strains, the P. putida KT2440 genomic database serves as...

The enigma of prokaryotic life in deep hypersaline anoxic basins (2005)

Bolhuis, H., Borin, S., Daffonchio, D., Corselli, C., Giuliano, L., ...

Deep hypersaline anoxic basins in the Mediterranean Sea are a legacy of dissolution of ancient subterranean salt deposits from the Miocene period. Our study revealed that these hypersaline basins are...

Substrate Specificity and Expression of Three 2,3-Dihydroxybiphenyl 1,2-Dioxygenases from Rhodococcus globerulus Strain P6 (2003)

McKay, D B, Prucha, M, Reineke, W, Timmis, K N, Pieper, D H

Rhodococcus globerulus strain P6 contains at least three genes, bphC1, bphC2, and bphC3, coding for 2,3-dihydroxybiphenyl 1,2-dioxygenases; the latter two specify enzymes of the family of one-domain...

Upstream binding sequences of the XylR activator protein and integration host factor in the xylS gene promoter region of the Pseudomonas TOL plasmid (1992)

Holtel, A., Timmis, K. N., Ramos, Juan L.

Final full-text version of the paper available at: http://nar.oxfordjournals.org/cgi/content/abstract/20/7/1755 .-- Copyright © by Oxford University Press. -- Http://nar.oxfordjournals.org/

A family of positive regulators related to the Pseudomonas putida TOL plasmid XylS and the Escherichia coli AraC activators (1990)

Ramos, Juan L., Rojo, F., Zhou, L., Timmis, K. N.

Final full-text version of the paper available at: http://nar.oxfordjournals.org/cgi/content/abstract/18/8/2149 .-- Copyright © by Oxford University Press. -- Http://nar.oxfordjournals.org/

A family of positive regulators related to the Pseudomonas putida TOL plasmid XylS and the Escherichia coli AraC activators (1990)

Ramos, J.L., Rojo, F., Zhou, L., Timmis, K.N.

The XylS family consists of at least 8 different transcriptional regulators. Six of these proteins are positive regulators for the catabolism of carbon sources (benzoate and sugars) in Escherichia...

Removal of Mercury from Chloralkali Electrolysis Wastewater by a Mercury-Resistant Pseudomonas putida Strain

Von Canstein, H., Li, Y., Timmis, K. N., Wagner-Döbler, I.

A mercury-resistant bacterial strain which is able to reduce ionic mercury to metallic mercury was used to remediate in laboratory columns mercury-containing wastewater produced during electrolytic...

Characterization of Three XylT-Like [2Fe-2S] Ferredoxins Associated with Catabolism of Cresols or Naphthalene: Evidence for Their Involvement in Catechol Dioxygenase Reactivation

Hugo, N., Meyer, C., Armengaud, J., Gaillard, J., Timmis, K. N., Jouanneau, Y.

The xylT gene product, a component of the xylene catabolic pathway of Pseudomonas putida mt2, has been recently characterized as a novel [2Fe-2S] ferredoxin which specifically reactivates...

Characterization of a new lipopeptide surfactant produced by thermotolerant and halotolerant subsurface Bacillus licheniformis BAS50.

Yakimov, M M, Timmis, K N, Wray, V, Fredrickson, H L

Strain BAS50, isolated from a petroleum reservoir at a depth of 1,500 m and identified as Bacillus licheniformis, grew and produced a lipopeptide surfactant when cultured on a variety of substrates...

Conversion of chlorobiphenyls into phenylhexadienoates and benzoates by the enzymes of the upper pathway for polychlorobiphenyl degradation encoded by the bph locus of Pseudomonas sp. strain LB400.

Seeger, M, Timmis, K N, Hofer, B

Metabolism of 21 chlorobiphenyls by the enzymes of the upper biphenyl catabolic pathway encoded by the bph locus of Pseudomonas sp. strain LB400 was investigated by using recombinant strains...

Stable expression of pertussis toxin in Bordetella bronchiseptica under the control of a tightly regulated promoter.

Suarez, A, Staendner, L H, Rohde, M, Piatti, G, Timmis, K N, Guzmán, C A

Pertussis toxin (PT) is an essential component of accellular vaccines against whooping cough. However, the industrial production of PT from Bordetella pertussis is impaired by slow growth and poor...

Interaction of Listeria monocytogenes with mouse dendritic cells.

Guzman, C A, Rohde, M, Chakraborty, T, Domann, E, Hudel, M, Wehland, J, ...

In this study, the interaction of murine dendritic cells with Listeria monocytogenes was investigated. Dendritic cells are efficient antigen-presenting cells, play a key role in the immune response,...

Identification of an immunoglobulin A binding motif located in the beta-antigen of the c protein complex of group B streptococci.

Jerlström, P G, Talay, S R, Valentin-Weigand, P, Timmis, K N, Chhatwal, G S

The beta-antigen of the c protein complex of group B streptococci contains two immunoglobulin A (IgA)-binding domains called A and B. A 73-amino-acid segment in domain A is responsible for most of...

Construction and characterization of a live attenuated vaccine candidate against Shigella dysenteriae type 1.

Klee, S R, Tzschaschel, B D, Fält, I, Kärnell, A, Lindberg, A A, Timmis, K N, ...

Vaccine candidates against Shigella dysenteriae type 1, which is associated with the most severe cases of bacillary dysentery, were constructed. The rfp and rfb gene clusters, which code for S....

Involvement of IHF protein in expression of the Ps promoter of the Pseudomonas putida TOL plasmid.

Holtel, A, Goldenberg, D, Giladi, H, Oppenheim, A B, Timmis, K N

Regulation of the xyl gene operons of the Pseudomonas putida TOL plasmid is mediated by the products of the downstream clustered and divergently oriented xylR and xylS regulatory genes. The xylR-xylS...

Characterization of the basic replicon of pCM1, a narrow-host-range plasmid from the moderate halophile Chromohalobacter marismortui.

Mellado, E, Asturias, J A, Nieto, J J, Timmis, K N, Ventosa, A

The moderately halophilic bacterium Chromohalobacter marismortui contains a 17.5-kb narrow-host-range plasmid, pCM1, which shows interesting properties for the development of cloning vectors for the...

Expression of Shigella dysenteriae serotype 1 O-antigenic polysaccharide by Shigella flexneri aroD vaccine candidates and different S. flexneri serotypes.

Fält, I C, Schweda, E K, Klee, S, Singh, M, Floderus, E, Timmis, K N, ...

The potential utility of Shigella flexneri aroD vaccine candidates for the development of bi- or multivalent vaccines has been explored by the introduction of the genetic determinants rfp and rfb for...

Distance between alleles as a determinant of linkage in natural transformation of Acinetobacter calcoaceticus.

Kloos, D U, DiMarco, A A, Elsemore, D A, Timmis, K N, Ornston, L N

Cotransformation frequencies of 16, 39, 51, and 60% were observed when donor alleles were separated by distances of 9.2, 7.4, 6.3, and 5.1 kb, respectively, in donor Acinetobacter calcoaceticus DNA....

Metal selectivity of in situ microcolonies in biofilms of the Elbe river.

Lünsdorf, H, Brümmer, I, Timmis, K N, Wagner-Döbler, I

The ultrastructure of natural complex biofilm communities of the Elbe river grown in situ on microscopic glass coverslips was studied by using transmission electron microscopy and energy-dispersive...

Heterologous expression of biphenyl dioxygenase-encoding genes from a gram-positive broad-spectrum polychlorinated biphenyl degrader and characterization of chlorobiphenyl oxidation by the gene products.

McKay, D B, Seeger, M, Zielinski, M, Hofer, B, Timmis, K N

The bphA1A2A3A4 gene cluster, encoding a biphenyl dioxygenase from Rhodococcus globerulus P6, a gram-positive microorganism able to degrade a wide spectrum of polychlorobiphenyls (PCBs), was...

Influence of different rol gene products on the chain length of Shigella dysenteriae type 1 lipopolysaccharide O antigen expressed by Shigella flexneri carrier strains.

Klee, S R, Tzschaschel, B D, Timmis, K N, Guzman, C A

Introduction of the rol genes of Shigella dysenteriae 1 and Escherichia coli K-12 into Shigella flexneri carrier strains expressing the heterologous S. dysenteriae type 1 lipopolysaccharide resulted...

Degradation of trichloroethylene by Pseudomonas cepacia G4 and the constitutive mutant strain G4 5223 PR1 in aquifer microcosms.

Krumme, M L, Timmis, K N, Dwyer, D F

Pseudomonas cepacia G4 degrades trichloroethylene (TCE) via a degradation pathway for aromatic compounds which is induced by substrates such as phenol and tryptophan. P. cepacia G4 5223 PR1 (PR1) is...

Biotransformation of substituted benzoates to the corresponding cis-diols by an engineered strain of Pseudomonas oleovorans producing the TOL plasmid-specified enzyme toluate-1,2-dioxygenase.

Wubbolts, M G, Timmis, K N

The conversion of substituted benzoates into 1,2-cis-dihydroxycyclohexa-3,5-diene carboxylic acids (cis-diols) was effected by using Escherichia coli and Pseudomonas recombinants carrying the xylXYZ...

Two independently regulated cytochromes P-450 in a Rhodococcus rhodochrous strain that degrades 2-ethoxyphenol and 4-methoxybenzoate.

Karlson, U, Dwyer, D F, Hooper, S W, Moore, E R, Timmis, K N, Eltis, L D

A red-pigmented coryneform bacterium, identified as Rhodococcus rhodochrous strain 116, that grew on 2-ethoxyphenol and 4-methoxybenzoate as sole carbon and energy sources was isolated. Phylogenetic...

Evaluation of aquatic sediment microcosms and their use in assessing possible effects of introduced microorganisms on ecosystem parameters.

Wagner-Döbler, I, Pipke, R, Timmis, K N, Dwyer, D F

In this paper we describe a sediment microcosm system consisting of 20 undisturbed, layered sediment cores with overlying site water which are incubated under identical conditions of temperature,...

Survival and function of a genetically engineered Pseudomonad in aquatic sediment microcosms.

Pipke, R, Wagner-Döbler, I, Timmis, K N, Dwyer, D F

Pseudomonas sp. strain B13 FR1(pFRC20P) is a genetically engineered microorganism (GEM) which is able to degrade chloro- and methylaromatics through a constructed ortho cleavage pathway. The fate of...

Cross talk between catabolic pathways in Pseudomonas putida: XylS-dependent and -independent activation of the TOL meta operon requires the same cis-acting sequences within the Pm promoter.

Kessler, B, Marqués, S, Köhler, T, Ramos, J L, Timmis, K N, De Lorenzo, V

The Pm promoter of the meta cleavage operon in the TOL (toluene degradation) plasmid pWW0 of Pseudomonas putida becomes activated by the plasmid-encoded XylS regulator in the presence of benzoate and...

Substrate specificity of catechol 2,3-dioxygenase encoded by TOL plasmid pWW0 of Pseudomonas putida and its relationship to cell growth.

Cerdan, P, Wasserfallen, A, Rekik, M, Timmis, K N, Harayama, S

Catechol 2,3-dioxygenase encoded by TOL plasmid pWW0 of Pseudomonas putida consists of four identical subunits, each containing one ferrous ion. The enzyme catalyzes ring cleavage of catechol,...

Inducible cell lysis system for the study of natural transformation and environmental fate of DNA released by cell death.

Kloos, D U, Strätz, M, Güttler, A, Steffan, R J, Timmis, K N

Two novel conditional broad-host-range cell lysis systems have been developed for the study of natural transformation in bacteria and the environmental fate of DNA released by cell death. Plasmid...

Influence of para-substituents on the oxidative metabolism of o-nitrophenols by Pseudomonas putida B2.

Zeyer, J, Kocher, H P, Timmis, K N

Pseudomonas putida B2 is able to grow on o-nitrophenol (ONP) as the sole source of carbon and nitrogen. ONP was converted by a nitrophenol oxygenase to nitrite and catechol. Catechol was then...

Relatedness of a periplasmic, broad-specificity RNase from Aeromonas hydrophila to RNase I of Escherichia coli and to a family of eukaryotic RNases.

Favre, D, Ngai, P K, Timmis, K N

The isolation, sequencing, and characterization of a periplasmic RNase gene from Aeromonas hydrophila AH1133 is described. Following subcloning of the gene on a 2.7-kb PstI fragment, its direction of...

Three different 2,3-dihydroxybiphenyl-1,2-dioxygenase genes in the gram-positive polychlorobiphenyl-degrading bacterium Rhodococcus globerulus P6.

Asturias, J A, Timmis, K N

Rhodococcus globerulus P6 (previously designated Acinetobacter sp. strain P6, Arthrobacter sp. strain M5, and Corynebacterium sp. strain MB1) is able to degrade a wide range of polychlorinated...

Carbon source-dependent inhibition of xyl operon expression of the Pseudomonas putida TOL plasmid.

Holtel, A, Marqués, S, Möhler, I, Jakubzik, U, Timmis, K N

TOL plasmid-encoded degradation of benzyl alcohol by Pseudomonas putida is inhibited by glucose and other compounds related to the main carbohydrate metabolism in Pseudomonas species. We report here...

Transcriptional induction kinetics from the promoters of the catabolic pathways of TOL plasmid pWW0 of Pseudomonas putida for metabolism of aromatics.

Marqués, S, Holtel, A, Timmis, K N, Ramos, J L

We determined, under several growth conditions, the kinetics of mRNA synthesis from the four Pseudomonas putida pWW0 plasmid promoters involved in degradation of xylenes and methylbenzyl alcohols via...

Genetic evidence that the XylS regulator of the Pseudomonas TOL meta operon controls the Pm promoter through weak DNA-protein interactions.

Kessler, B, Herrero, M, Timmis, K N, De Lorenzo, V

The activation of the Pm promoter of the meta operon of the TOL plasmid of Pseudomonas putida by its cognate XylS activator protein in the presence and absence of benzoate inducers has been examined...

Early and late responses of TOL promoters to pathway inducers: identification of postexponential promoters in Pseudomonas putida with lacZ-tet bicistronic reporters.

De Lorenzo, V, Cases, I, Herrero, M, Timmis, K N

Transcriptional lacZ fusions to the Pu and Pm promoters of the TOL (toluene degradation) plasmid inserted in monocopy in the chromosome of Pseudomonas putida showed a very different responsiveness to...

Characterization of 2,2',3-trihydroxybiphenyl dioxygenase, an extradiol dioxygenase from the dibenzofuran- and dibenzo-p-dioxin-degrading bacterium Sphingomonas sp. strain RW1.

Happe, B, Eltis, L D, Poth, H, Hedderich, R, Timmis, K N

A key enzyme in the degradation pathways of dibenzo-p-dioxin and dibenzofuran, namely, 2,2',3-trihydroxybiphenyl dioxygenase, which is responsible for meta cleavage of the first aromatic ring, has...

Involvement of Pseudomonas putida RpoN sigma factor in regulation of various metabolic functions.

Köhler, T, Harayama, S, Ramos, J L, Timmis, K N

The RpoN protein was originally identified in Escherichia coli as a sigma (sigma) factor essential for the expression of nitrogen regulons. In the present study we cloned the Pseudomonas putida rpoN...

Characterization of five genes in the upper-pathway operon of TOL plasmid pWW0 from Pseudomonas putida and identification of the gene products.

Harayama, S, Rekik, M, Wubbolts, M, Rose, K, Leppik, R A, Timmis, K N

The upper operon of the TOL plasmid pWW0 of Pseudomonas putida encodes a set of enzymes which transform toluene and xylenes to benzoate and toluates. The genetic organization of the operon was...

Regulator and enzyme specificities of the TOL plasmid-encoded upper pathway for degradation of aromatic hydrocarbons and expansion of the substrate range of the pathway.

Abril, M A, Michan, C, Timmis, K N, Ramos, J L

The TOL plasmid upper pathway operon encodes enzymes involved in the catabolism of aromatic hydrocarbons such as toluene and xylenes. The regulator of the gene pathway, the XylR protein, exhibits a...

Roles of the divergent branches of the meta-cleavage pathway in the degradation of benzoate and substituted benzoates.

Harayama, S, Mermod, N, Rekik, M, Lehrbach, P R, Timmis, K N

The TOL plasmid-specified meta-cleavage pathway for the oxidative catabolism of benzoate and toluates branches at the ring cleavage products of catechols and reconverges later at 2-oxopent-4-enoate...

Analysis, cloning, and high-level expression of 2,4-dichlorophenoxyacetate monooxygenase gene tfdA of Alcaligenes eutrophus JMP134.

Streber, W R, Timmis, K N, Zenk, M H

Plasmid pJP4 of Alcaligenes eutrophus JMP134 contains all genes for the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D). Five of these genes, tfdB, tfdC, tfdD, tfdE, and tfdF, have recently...

Vector for regulated expression of cloned genes in a wide range of gram-negative bacteria.

Mermod, N, Ramos, J L, Lehrbach, P R, Timmis, K N

A pKT231-based broad-host-range plasmid vector was constructed which enabled regulation of expression of cloned genes in a wide range of gram-negative bacteria. This vector, pNM185, contained...

Gene order of the TOL catabolic plasmid upper pathway operon and oxidation of both toluene and benzyl alcohol by the xylA product.

Harayama, S, Leppik, R A, Rekik, M, Mermod, N, Lehrbach, P R, Reineke, W, ...

TOL plasmid pWW0 specifies enzymes for the oxidative catabolism of toluene and xylenes. The upper pathway converts the aromatic hydrocarbons to aromatic carboxylic acids via corresponding alcohols...

Mutations leading to constitutive expression from the TOL plasmid meta-cleavage pathway operon are located at the C-terminal end of the positive regulator protein XylS.

Zhou, L M, Timmis, K N, Ramos, J L

The XylS protein is the positive activator of the TOL plasmid meta-cleavage pathway operon for the metabolism of alkylbenzoates in Pseudomonas putida. The regulator stimulates transcription from the...

Characterization of a plasmid-specified pathway for catabolism of isopropylbenzene in Pseudomonas putida RE204.

Eaton, R W, Timmis, K N

A Pseudomonas putida strain designated RE204, able to utilize isopropylbenzene as the sole carbon and energy source, was isolated. Tn5 transposon mutagenesis by means of the suicide transposon donor...

Spontaneous deletion of a 20-kilobase DNA segment carrying genes specifying isopropylbenzene metabolism in Pseudomonas putida RE204.

Eaton, R W, Timmis, K N

The genes encoding isopropylbenzene metabolism in Pseudomonas putida RE204 are readily lost in two ways: by loss (curing) of plasmid pRE4 which specifies the catabolic pathway and by deletion from...

Highly repressible expression system for cloning genes that specify potentially toxic proteins.

O'Connor, C D, Timmis, K N

A highly repressible expression vector system that allows the cloning of potentially deleterious genes has been constructed. Undesired expression of a cloned gene was prevented (i) at the level of...

Transposon mutagenesis analysis of meta-cleavage pathway operon genes of the TOL plasmid of Pseudomonas putida mt-2.

Harayama, S, Lehrbach, P R, Timmis, K N

Hybrid plasmids containing the regulated meta-cleavage pathway operon of TOL plasmid pWWO were mutagenized with transposon Tn1000 or Tn5. The resulting insertion mutant plasmids were examined for...

Transposon mutagenesis and cloning analysis of the pathways for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP134(pJP4).

Don, R H, Weightman, A J, Knackmuss, H J, Timmis, K N

Plasmid pJP4 permits its host bacterium, strain JMP134, to degrade and utilize as sole sources of carbon and energy 3-chlorobenzoate and 2,4-dichlorophenoxyacetic acid (R. H. Don and J. M. Pemberton,...

Enzyme recruitment in vitro: use of cloned genes to extend the range of haloaromatics degraded by Pseudomonas sp. strain B13.

Lehrbach, P R, Zeyer, J, Reineke, W, Knackmuss, H J, Timmis, K N

DNA fragments containing the xylD and xylL genes of TOL plasmid pWW0 -161 of Pseudomonas putida, which code for the catabolic enzymes toluate 1,2-dioxygenase and dihydrodihydroxybenzoic acid...

Replication control mutations of plasmid R6-5 and their effects on interactions of the RNA-I control element with its target.

Brady, G, Frey, J, Danbara, H, Timmis, K N

Nine high copy number mutations of plasmid R6-5, representing five phenotypically distinct groups, have been identified by DNA sequencing. In each mutant plasmid examined, a single nucleotide change...

Localization and functional analysis of transposon mutations in regulatory genes of the TOL catabolic pathway.

Franklin, F C, Lehrbach, P R, Lurz, R, Rueckert, B, Bagdasarian, M, Timmis, K N

Mutant derivatives of the TOL plasmid pWW0-161, containing Tn5 insertions in the xylS and xylR regulatory genes of the catabolic pathway, have been identified and characterized. The two genes are...

Microbial mineralization of ring-substituted anilines through an ortho-cleavage pathway.

Zeyer, J, Wasserfallen, A, Timmis, K N

Moraxella sp. strain G is able to utilize as sole source of carbon and nitrogen aniline, 4-fluoroaniline, 2-chloroaniline, 3-chloroaniline, 4-chloroaniline (PCA), and 4-bromoaniline but not...

Use of cloned genes of Pseudomonas TOL plasmid to effect biotransformation of benzoates to cis-dihydrodiols and catechols by Escherichia coli cells.

Zeyer, J, Lehrbach, P R, Timmis, K N

DNA fragments containing the xylD and xylL genes, which specify the broad-specificity enzymes toluate-1,2-dioxygenase and 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid dehydrogenase, respectively, of...

Construction of stable LamB-Shiga toxin B subunit hybrids: analysis of expression in Salmonella typhimurium aroA strains and stimulation of B subunit-specific mucosal and serum antibody responses.

Su, G F, Brahmbhatt, H N, Wehland, J, Rohde, M, Timmis, K N

The complete Shiga toxin B subunit and two N-terminal segments of the B subunit have been inserted into a cell surface exposed loop of the LamB protein, and expression of the hybrid proteins from...

Fibronectin-binding protein of Streptococcus pyogenes: sequence of the binding domain involved in adherence of streptococci to epithelial cells.

Talay, S R, Valentin-Weigand, P, Jerlström, P G, Timmis, K N, Chhatwal, G S

The sequence of the fibronectin-binding domain of the fibronectin-binding protein of Streptococcus pyogenes (Sfb protein) was determined, and its role in streptococcal adherence was investigated by...

Specific lung mucosal and systemic immune responses after oral immunization of mice with Salmonella typhimurium aroA, Salmonella typhi Ty21a, and invasive Escherichia coli expressing recombinant pertussis toxin S1 subunit.

Walker, M J, Rohde, M, Timmis, K N, Guzmán, C A

Pertussis toxin (PT) is considered an essential protective component for incorporation into new generation vaccines against Bordetella pertussis, the causative agent of whooping cough. Traditionally,...

Production of recombinant Bordetella pertussis serotype 2 fimbriae in Bordetella parapertussis and Bordetella bronchiseptica: utility of Escherichia coli gene expression signals.

Walker, M J, Guzmán, C A, Rohde, M, Timmis, K N

Serotype-specific fimbriae of Bordetella pertussis are considered potential components of new-generation vaccines against whooping cough. Attempts to characterize fimbriae, and indeed other virulence...

Intercellular spread of Shigella flexneri through a monolayer mediated by membranous protrusions and associated with reorganization of the cytoskeletal protein vinculin.

Kadurugamuwa, J L, Rohde, M, Wehland, J, Timmis, K N

The spread of Shigella flexneri in a monolayer of infected Henle and HeLa cells was studied by using immunofluorescence and electron microscopy. Infected cells produced numerous bacterium-containing...

Direct expression of Bordetella pertussis filamentous hemagglutinin in Escherichia coli and Salmonella typhimurium aroA.

Guzmán, C A, Walker, M J, Rohde, M, Timmis, K N

Nonfused (i.e., nonhybrid) filamentous hemagglutinin (FHA) of Bordetella pertussis was efficiently expressed in Escherichia coli K-12 and Salmonella typhimurium aroA at levels higher than those found...

Construction of minitransposons for constitutive and inducible expression of pertussis toxin in bvg-negative Bordetella bronchiseptica.

Walker, M J, Rohde, M, Wehland, J, Timmis, K N

Appropriately detoxified pertussis toxin (PT) of Bordetella pertussis is considered to be an essential component of new-generation whooping cough vaccines, but the development of a procedure to...

Characterization of murine monoclonal antibodies that recognize defined epitopes of pertussis toxin and neutralize its toxic effect on Chinese hamster ovary cells.

Walker, M J, Wehland, J, Timmis, K N, Raupach, B, Schmidt, M A

Three murine monoclonal antibodies (MAb), E19, E205, and E251, raised against pertussis toxin reacted in Western blots (immunoblots) with the S1, S4, and S2-S3 subunits, respectively, and neutralized...

Antibody responses in the lungs of mice following oral immunization with Salmonella typhimurium aroA and invasive Escherichia coli strains expressing the filamentous hemagglutinin of Bordetella pertussis.

Guzmán, C A, Brownlie, R M, Kadurugamuwa, J, Walker, M J, Timmis, K N

The filamentous hemagglutinin (FHA) of Bordetella pertussis was expressed in the attenuated aroA mutant of Salmonella typhimurium, SL3261, and in a strain of Escherichia coli harboring Shigella...

Cloning and expression in Escherichia coli of the Ibc protein genes of group B streptococci: binding of human immunoglobulin A to the beta antigen.

Cleat, P H, Timmis, K N

Total-cell DNA isolated from a highly virulent serotype Ic strain of a group B streptococcus was used to construct a gene bank with bacteriophage lambda L47.1 in Escherichia coli K-12. Recombinant...

Localization of stx, a determinant essential for high-level production of shiga toxin by Shigella dysenteriae serotype 1, near pyrF and generation of stx transposon mutants.

Sekizaki, T, Harayama, S, Brazil, G M, Timmis, K N

Hfr strains of Shigella dysenteriae serotype 1 were constructed by transient integration of an RP4 plasmid derivative carrying transposon Tn501 into the Shigella chromosome through Tn501-mediated...

Small virulence plasmid of Shigella dysenteriae 1 strain W30864 encodes a 41,000-dalton protein involved in formation of specific lipopolysaccharide side chains of serotype 1 isolates.

Watanabe, H, Nakamura, A, Timmis, K N

A 6-megadalton plasmid, pHW400, of Shigella dysenteriae 1 strain W30864 was previously found to specify one or more functions for O-antigen production and bacterial virulence (H. Watanabe and K. N....

Monoclonal antibody detection of IncF group plasmid-encoded TraT protein in clinical isolates of Escherichia coli.

Bitter-Suermann, D, Peters, H, Jürs, M, Nehrbass, R, Montenegro, M, Timmis, K N

The TraT protein specified by IncF group plasmids mediates surface exclusion and bacterial resistance to the lethal activities of serum. In this study, an anti-TraT protein monoclonal antibody was...

Cloning of enterotoxin gene from Aeromonas hydrophila provides conclusive evidence of production of a cytotonic enterotoxin.

Chakraborty, T, Montenegro, M A, Sanyal, S C, Helmuth, R, Bulling, E, Timmis, K N

Culture filtrates of two Aeromonas hydrophila strains which were isolated from patients with diarrhea and assumed to be causative agents of the infections were shown to contain enterotoxic,...

A plasmid-encoded outer membrane protein, TraT, enhances resistance of Escherichia coli to phagocytosis.

Agüero, M E, Aron, L, DeLuca, A G, Timmis, K N, Cabello, F C

The presence of the outer membrane protein TraT, encoded by plasmid R6-5, reduces the sensitivity of Escherichia coli cells to phagocytosis by macrophages. This effect is independent of the bacterial...

Cloning and genetic analysis of serotype 5 M protein determinant of group A streptococci: evidence for multiple copies of the M5 determinant in the Streptococcus pyogenes genome.

Kehoe, M A, Poirier, T P, Beachey, E H, Timmis, K N

A gene bank of group A Streptococcus strain Manfredo (M protein serotype 5) was constructed with a bacteriophage lambda vector-Escherichia coli K-12 host system and screened by immunoblotting hybrid...

Expression of protective and cardiac tissue cross-reactive epitopes of type 5 streptococcal M protein in Escherichia coli.

Poirier, T P, Kehoe, M A, Dale, J B, Timmis, K N, Beachey, E H

The immunochemical properties of type 5 M protein antigens that were expressed in Escherichia coli K-12 by recombinant lambda bacteriophages isolated from a gene bank of serotype 5 Streptococcus...

Monoclonal antibodies to enterobacterial common antigen and to Escherichia coli lipopolysaccharide outer core: demonstration of an antigenic determinant shared by enterobacterial common antigen and E. coli K5 capsular polysaccharide.

Peters, H, Jürs, M, Jann, B, Jann, K, Timmis, K N, Bitter-Suermann, D

We established hybridoma cell lines producing monoclonal antibodies against enterobacterial common antigen (ECA) and a substructure of the outer core of different Escherichia coli lipopolysaccharides...

A small plasmid in Shigella dysenteriae 1 specifies one or more functions essential for O antigen production and bacterial virulence.

Watanabe, H, Timmis, K N

The role of plasmids in the virulence of Shigella dysenteriae 1 W30864, which contains at least five species, was investigated. By means of a standard plasmid-curing procedure, that is, bacterial...

Cloning and expression in Escherichia coli of the streptolysin O determinant from Streptococcus pyogenes: characterization of the cloned streptolysin O determinant and demonstration of the absence of substantial homology with determinants of other thiol-activated toxins.

Kehoe, M, Timmis, K N

A gene bank of Streptococcus pyogenes Richards was constructed in Escherichia coli by using the bacteriophage replacement vector lambda L47.1, and hybrid phage expressing streptolysin O (SLO) were...

Cloning and analysis of the K1 capsule biosynthesis genes of Escherichia coli: lack of homology with Neisseria meningitidis group B DNA sequences.

Echarti, C, Hirschel, B, Boulnois, G J, Varley, J M, Waldvogel, F, Timmis, K N

Genes coding for production of the K1 polysaccharide capsule of Escherichia coli have been cloned. Complementation, insertion, and deletion analyses were used to localize the K1 genes and...

Location of two relaxation nick sites in R6K and single sites in pSC101 and RSF1010 close to origins of vegetative replication: implication for conjugal transfer of plasmid deoxyribonucleic acid.

Nordheim, A, Hashimoto-Gotoh, T, Timmis, K N

A nick-labeling method has been used to localize the relaxation complex nick sites in three plasmids (pSC101, RSF1010, and R6K) that differ markedly in their host range, deoxyribonucleic acid...

Plasmid replication functions: two distinct segments of plasmid R1, RepA and RepD, express incompatibility and are capable of autonomous replication.

Danbara, H, Timmis, J K, Lurz, R, Timmis, K N

The genetic determinants for replication and incompatibility of plasmid R1 were investigated by gene cloning methods, and three types of R1 miniplasmid derivatives were generated. The first,...

Antibody responses in the serum and respiratory tract of mice following oral vaccination with liposomes coated with filamentous hemagglutinin and pertussis toxoid.

Guzman, C A, Molinari, G, Fountain, M W, Rohde, M, Timmis, K N, Walker, M J

Mice were orally vaccinated with liposomes coated with filamentous hemagglutinin (FHA) and detoxified pertussis toxin (PT) of Bordetella pertussis. FHA- and PT-specific immunoglobulin G (IgG) was...

Invasion and intracellular survival of Bordetella bronchiseptica in mouse dendritic cells.

Guzman, C A, Rohde, M, Bock, M, Timmis, K N

We have studied the interaction between the respiratory pathogen Bordetella bronchiseptica and murine spleen dendritic cells, important antigen-presenting cells that are found in the airway...

Mechanisms involved in uptake of Bordetella bronchiseptica by mouse dendritic cells.

Guzman, C A, Rohde, M, Timmis, K N

The invasion and intracellular survival of Bordetella bronchiseptica in mouse dendritic cells were investigated. The results obtained suggest that B. bronchiseptica binds specifically to glycosylated...

Metagenome Survey of Biofilms in Drinking-Water Networks

Schmeisser, C., Stöckigt, C., Raasch, C., Wingender, J., Timmis, K. N., Wenderoth, D. F., ...

Most naturally occurring biofilms contain a vast majority of microorganisms which have not yet been cultured, and therefore we have little information on the genetic information content of these...

Upstream binding sequences of the XylR activator protein and integration host factor in the xylS gene promoter region of the Pseudomonas TOL plasmid.

Holtel, A, Timmis, K N, Ramos, J L

The xylR and xylS genes, which encode the positive regulators of the TOL plasmid catabolic pathways, are adjacent genes on the TOL plasmid and are transcribed from divergent promoters. Transcription...

Bacterial infection of wounds: fibronectin-mediated adherence group A and C streptococci to fibrin thrombi in vitro.

Chhatwal, G S, Valentin-Weigand, P, Timmis, K N

Adherence of group A, B, and C streptococci to fibrin thrombi was studied by using a novel fluorochrome microassay carried out in microdilution plates in which fibrin thrombi had been prepared by...

Regulation of DNA replication: "target" determinant of the replication control elements of plasmid R6-5 lies within a control element gene.

Danbara, H, Brady, G, Timmis, J K, Timmis, K N

The replication control system of plasmid R6-5 has been investigated by characterization of high-copy-number mutant miniplasmids, development of an in vivo assay for the site of action or "target" of...

A family of positive regulators related to the Pseudomonas putida TOL plasmid XylS and the Escherichia coli AraC activators.

Ramos, J L, Rojo, F, Zhou, L, Timmis, K N

The XylS family consists of a least 8 different transcriptional regulators. Six of these proteins are positive regulators for the catabolism of carbon sources (benzoate and sugars) in Escherichia...

Cloning of chicken lysozyme structural gene sequences synthesized in vitro.

Sippel, A E, Land, H, Lindenmaier, W, Nguyen-Huu, M C, Wurtz, T, Timmis, K N, ...

Double-stranded chicken lysozyme cDNA was synthesized from an oviduct mRNA fraction enriched for lysozyme mRNA. The ds-cDNA was inserted into the BamHI site of plasmid pBR322 using chemically...

Plasmid replication functions. IV. Promoters in the replication region of plasmid R6-5.

Slocombe, P M, Ely, S, Timmis, K N

Eight RNA polymerase binding sites have been shown to map within the EcoRI fragment E-2 (replication region RepA EcoRI fragment) of plasmid R6-5 and all but one have been shown to contain active...

Molecular and functional analysis of the TOL plasmid pWWO from Pseudomonas putida and cloning of genes for the entire regulated aromatic ring meta cleavage pathway.

Franklin, F C, Bagdasarian, M, Bagdasarian, M M, Timmis, K N

The genetic organization of the Pseudomonas putida plasmid pWWO-161, which encodes enzymes for the degradation of toluene and related aromatic hydrocarbons, has been investigated by transposition...

An inhibitor of SOS induction, specified by a plasmid locus in Escherichia coli.

Bagdasarian, M, Bailone, A, Bagdasarian, M M, Manning, P A, Lurz, R, Timmis, K N, ...

Plasmid R6-5 contains a locus whose product inhibits induction of sfiA and prophage lambda in a recA441 mutant at 42 degrees C and in a recA+ host after treatment with nalidixic acid. This plasmidic...

Altered effector specificities in regulators of gene expression: TOL plasmid xylS mutants and their use to engineer expansion of the range of aromatics degraded by bacteria.

Ramos, J L, Stolz, A, Reineke, W, Timmis, K N

Stimulation of transcription from positively regulated promoters involves regulatory proteins that have been activated, generally as a consequence of binding low molecular weight effector molecules....

NZB mouse system for production of monoclonal antibodies to weak bacterial antigens: isolation of an IgG antibody to the polysaccharide capsules of Escherichia coli K1 and group B meningococci.

Frosch, M, Görgen, I, Boulnois, G J, Timmis, K N, Bitter-Suermann, D

A system for the production of monoclonal antibodies, particularly of the IgG type, against weakly immunogenic bacterial polysaccharide antigens is described. This system, which is based on the...

An upstream XylR- and IHF-induced nucleoprotein complex regulates the sigma 54-dependent Pu promoter of TOL plasmid.

De Lorenzo, V, Herrero, M, Metzke, M, Timmis, K N

Transcription from promoter Pu of the upper catabolic operon of the Pseudomonas putida TOL plasmid which specifies conversion of toluene/xylenes to benzoate/toluates is activated by the TOL-encoded...

Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria.

Herrero, M, De Lorenzo, V, Timmis, K N

A simple procedure for cloning and stable insertion of foreign genes into the chromosomes of gram-negative eubacteria was developed by combining in two sets of plasmids (i) the transposition features...

Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria.

De Lorenzo, V, Herrero, M, Jakubzik, U, Timmis, K N

A collection of Tn5-derived minitransposons has been constructed that simplifies substantially the generation of insertion mutants, in vivo fusions with reporter genes, and the introduction of...

Plasmid-determined resistance to serum bactericidal activity: a major outer membrane protein, the traT gene product, is responsible for plasmid-specified serum resistance in Escherichia coli.

Moll, A, Manning, P A, Timmis, K N

Resistance to the bactericidal activity of serum appears to be an important virulence property of invasive bacteria. The conjugative multiple-antibiotic-resistance plasmid R6-5 was found to confer...

Transcription of the TOL plasmid toluate catabolic pathway operon of Pseudomonas putida is determined by a pair of co-ordinately and positively regulated overlapping promoters.

Mermod, N, Lehrbach, P R, Reineke, W, Timmis, K N

Expression of the meta-cleavage pathway operon of TOL plasmid pWW0 of Pseudomonas putida is positively regulated by the xylS gene product. We have sequenced the promoter region of this operon and...

Tracking the Response of Burkholderia cepacia G4 5223-PR1 in Aquifer Microcosms

Winkler, J., Timmis, K. N., Snyder, R. A.

The introduction of bacteria into the environment for bioremediation purposes (bioaugmentation) requires analysis and monitoring of microbial population dynamics to define persistence and activity...

Formation of Dimethylmuconolactones from Dimethylphenols by Alcaligenes eutrophus JMP 134

Pieper, D. H., Stadler-Fritzsche, K., Knackmuss, H., Timmis, K. N.

2,3-, 2,4-, 2,5-, 3,4-, and 3,5-dimethylphenols were cometabolized by 2,4-dichlorophenoxyacetate-grown Alcaligenes eutrophus JMP 134 or the constitutive derivative JMP 134-1 via the ortho pathway...

Degradation of Chlorinated Dibenzofurans and Dibenzo-p-Dioxins by Sphingomonas sp. Strain RW1

Wilkes, H., Wittich, R., Timmis, K. N., Fortnagel, P., Francke, W.

The ability of the dibenzofuran- and dibenzo-p-dioxin-mineralizing bacterium Sphingomonas sp. strain RW1 (R.-M. Wittich, H. Wilkes, V. Sinnwell, W. Francke, and P. Fortnagel, Appl. Environ....

Use of Colicin E3 for Biological Containment of Microorganisms

Munthali, M. T., Timmis, K. N., Diaz, E.

The genetic determinant of the lethal antibiotic colicin E3 was cloned under the control of a tightly regulated promoter in the absence of the gene for its cognate inhibitor. Combination of this...

Metabolism of Chlorotoluenes by Burkholderia sp. Strain PS12 and Toluene Dioxygenase of Pseudomonas putida F1: Evidence for Monooxygenation by Toluene and Chlorobenzene Dioxygenases

Lehning, A., Fock, U., Wittich, R., Timmis, K. N., Pieper, D. H.

The degradation of toluene by Pseudomonas putida F1 and of chlorobenzenes by Burkholderia sp. strain PS12 is initiated by incorporation of dioxygen into the aromatic nucleus to form...

Mineralization of 4-Chlorodibenzofuran by a Consortium Consisting of Sphingomonas sp. Strain RW1 and Burkholderia sp. Strain JWS

Arfmann, H., Timmis, K. N., Wittich, R.

The dibenzofuran-degrading bacterium Sphingomonas sp. strain RW1 (R.-M. Wittich, H. Wilkes, V. Sinnwell, W. Francke, and P. Fortnagel, Appl. Environ. Microbiol. 58:1005-1010, 1992) attacks...

Evidence that Formation of Protoanemonin from Metabolites of 4-Chlorobiphenyl Degradation Negatively Affects the Survival of 4-Chlorobiphenyl-Cometabolizing Microorganisms

Blasco, R., Mallavarapu, M., Wittich, R., Timmis, K. N., Pieper, D. H.

A rapid decline in cell viability of different PCB-metabolizing organisms was observed in soil microcosms amended with 4-chlorobiphenyl. The toxic effect could not be attributed to 4-chlorobiphenyl...

Removal of Mercury from Chloralkali Electrolysis Wastewater by a Mercury-Resistant Pseudomonas putida Strain

Von Canstein, H., Li, Y., Timmis, K. N., Wagner-Döbler, I.

A mercury-resistant bacterial strain which is able to reduce ionic mercury to metallic mercury was used to remediate in laboratory columns mercury-containing wastewater produced during electrolytic...

Characterization of Three XylT-Like [2Fe-2S] Ferredoxins Associated with Catabolism of Cresols or Naphthalene: Evidence for Their Involvement in Catechol Dioxygenase Reactivation

Hugo, N., Meyer, C., Armengaud, J., Gaillard, J., Timmis, K. N., Jouanneau, Y.

The xylT gene product, a component of the xylene catabolic pathway of Pseudomonas putida mt2, has been recently characterized as a novel [2Fe-2S] ferredoxin which specifically reactivates...

Characterization of a new lipopeptide surfactant produced by thermotolerant and halotolerant subsurface Bacillus licheniformis BAS50.

Yakimov, M M, Timmis, K N, Wray, V, Fredrickson, H L

Strain BAS50, isolated from a petroleum reservoir at a depth of 1,500 m and identified as Bacillus licheniformis, grew and produced a lipopeptide surfactant when cultured on a variety of substrates...

Conversion of chlorobiphenyls into phenylhexadienoates and benzoates by the enzymes of the upper pathway for polychlorobiphenyl degradation encoded by the bph locus of Pseudomonas sp. strain LB400.

Seeger, M, Timmis, K N, Hofer, B

Metabolism of 21 chlorobiphenyls by the enzymes of the upper biphenyl catabolic pathway encoded by the bph locus of Pseudomonas sp. strain LB400 was investigated by using recombinant strains...

Stable expression of pertussis toxin in Bordetella bronchiseptica under the control of a tightly regulated promoter.

Suarez, A, Staendner, L H, Rohde, M, Piatti, G, Timmis, K N, Guzmán, C A

Pertussis toxin (PT) is an essential component of accellular vaccines against whooping cough. However, the industrial production of PT from Bordetella pertussis is impaired by slow growth and poor...

Interaction of Listeria monocytogenes with mouse dendritic cells.

Guzman, C A, Rohde, M, Chakraborty, T, Domann, E, Hudel, M, Wehland, J, ...

In this study, the interaction of murine dendritic cells with Listeria monocytogenes was investigated. Dendritic cells are efficient antigen-presenting cells, play a key role in the immune response,...

Identification of an immunoglobulin A binding motif located in the beta-antigen of the c protein complex of group B streptococci.

Jerlström, P G, Talay, S R, Valentin-Weigand, P, Timmis, K N, Chhatwal, G S

The beta-antigen of the c protein complex of group B streptococci contains two immunoglobulin A (IgA)-binding domains called A and B. A 73-amino-acid segment in domain A is responsible for most of...

Construction and characterization of a live attenuated vaccine candidate against Shigella dysenteriae type 1.

Klee, S R, Tzschaschel, B D, Fält, I, Kärnell, A, Lindberg, A A, Timmis, K N, ...

Vaccine candidates against Shigella dysenteriae type 1, which is associated with the most severe cases of bacillary dysentery, were constructed. The rfp and rfb gene clusters, which code for S....

Involvement of IHF protein in expression of the Ps promoter of the Pseudomonas putida TOL plasmid.

Holtel, A, Goldenberg, D, Giladi, H, Oppenheim, A B, Timmis, K N

Regulation of the xyl gene operons of the Pseudomonas putida TOL plasmid is mediated by the products of the downstream clustered and divergently oriented xylR and xylS regulatory genes. The xylR-xylS...

Characterization of the basic replicon of pCM1, a narrow-host-range plasmid from the moderate halophile Chromohalobacter marismortui.

Mellado, E, Asturias, J A, Nieto, J J, Timmis, K N, Ventosa, A

The moderately halophilic bacterium Chromohalobacter marismortui contains a 17.5-kb narrow-host-range plasmid, pCM1, which shows interesting properties for the development of cloning vectors for the...

Expression of Shigella dysenteriae serotype 1 O-antigenic polysaccharide by Shigella flexneri aroD vaccine candidates and different S. flexneri serotypes.

Fält, I C, Schweda, E K, Klee, S, Singh, M, Floderus, E, Timmis, K N, ...

The potential utility of Shigella flexneri aroD vaccine candidates for the development of bi- or multivalent vaccines has been explored by the introduction of the genetic determinants rfp and rfb for...

Distance between alleles as a determinant of linkage in natural transformation of Acinetobacter calcoaceticus.

Kloos, D U, DiMarco, A A, Elsemore, D A, Timmis, K N, Ornston, L N

Cotransformation frequencies of 16, 39, 51, and 60% were observed when donor alleles were separated by distances of 9.2, 7.4, 6.3, and 5.1 kb, respectively, in donor Acinetobacter calcoaceticus DNA....

Metal selectivity of in situ microcolonies in biofilms of the Elbe river.

Lünsdorf, H, Brümmer, I, Timmis, K N, Wagner-Döbler, I

The ultrastructure of natural complex biofilm communities of the Elbe river grown in situ on microscopic glass coverslips was studied by using transmission electron microscopy and energy-dispersive...

Heterologous expression of biphenyl dioxygenase-encoding genes from a gram-positive broad-spectrum polychlorinated biphenyl degrader and characterization of chlorobiphenyl oxidation by the gene products.

McKay, D B, Seeger, M, Zielinski, M, Hofer, B, Timmis, K N

The bphA1A2A3A4 gene cluster, encoding a biphenyl dioxygenase from Rhodococcus globerulus P6, a gram-positive microorganism able to degrade a wide spectrum of polychlorobiphenyls (PCBs), was...

Influence of different rol gene products on the chain length of Shigella dysenteriae type 1 lipopolysaccharide O antigen expressed by Shigella flexneri carrier strains.

Klee, S R, Tzschaschel, B D, Timmis, K N, Guzman, C A

Introduction of the rol genes of Shigella dysenteriae 1 and Escherichia coli K-12 into Shigella flexneri carrier strains expressing the heterologous S. dysenteriae type 1 lipopolysaccharide resulted...

Degradation of trichloroethylene by Pseudomonas cepacia G4 and the constitutive mutant strain G4 5223 PR1 in aquifer microcosms.

Krumme, M L, Timmis, K N, Dwyer, D F

Pseudomonas cepacia G4 degrades trichloroethylene (TCE) via a degradation pathway for aromatic compounds which is induced by substrates such as phenol and tryptophan. P. cepacia G4 5223 PR1 (PR1) is...

Biotransformation of substituted benzoates to the corresponding cis-diols by an engineered strain of Pseudomonas oleovorans producing the TOL plasmid-specified enzyme toluate-1,2-dioxygenase.

Wubbolts, M G, Timmis, K N

The conversion of substituted benzoates into 1,2-cis-dihydroxycyclohexa-3,5-diene carboxylic acids (cis-diols) was effected by using Escherichia coli and Pseudomonas recombinants carrying the xylXYZ...

Two independently regulated cytochromes P-450 in a Rhodococcus rhodochrous strain that degrades 2-ethoxyphenol and 4-methoxybenzoate.

Karlson, U, Dwyer, D F, Hooper, S W, Moore, E R, Timmis, K N, Eltis, L D

A red-pigmented coryneform bacterium, identified as Rhodococcus rhodochrous strain 116, that grew on 2-ethoxyphenol and 4-methoxybenzoate as sole carbon and energy sources was isolated. Phylogenetic...

Evaluation of aquatic sediment microcosms and their use in assessing possible effects of introduced microorganisms on ecosystem parameters.

Wagner-Döbler, I, Pipke, R, Timmis, K N, Dwyer, D F

In this paper we describe a sediment microcosm system consisting of 20 undisturbed, layered sediment cores with overlying site water which are incubated under identical conditions of temperature,...

Survival and function of a genetically engineered Pseudomonad in aquatic sediment microcosms.

Pipke, R, Wagner-Döbler, I, Timmis, K N, Dwyer, D F

Pseudomonas sp. strain B13 FR1(pFRC20P) is a genetically engineered microorganism (GEM) which is able to degrade chloro- and methylaromatics through a constructed ortho cleavage pathway. The fate of...

Cross talk between catabolic pathways in Pseudomonas putida: XylS-dependent and -independent activation of the TOL meta operon requires the same cis-acting sequences within the Pm promoter.

Kessler, B, Marqués, S, Köhler, T, Ramos, J L, Timmis, K N, De Lorenzo, V

The Pm promoter of the meta cleavage operon in the TOL (toluene degradation) plasmid pWW0 of Pseudomonas putida becomes activated by the plasmid-encoded XylS regulator in the presence of benzoate and...

Substrate specificity of catechol 2,3-dioxygenase encoded by TOL plasmid pWW0 of Pseudomonas putida and its relationship to cell growth.

Cerdan, P, Wasserfallen, A, Rekik, M, Timmis, K N, Harayama, S

Catechol 2,3-dioxygenase encoded by TOL plasmid pWW0 of Pseudomonas putida consists of four identical subunits, each containing one ferrous ion. The enzyme catalyzes ring cleavage of catechol,...

Inducible cell lysis system for the study of natural transformation and environmental fate of DNA released by cell death.

Kloos, D U, Strätz, M, Güttler, A, Steffan, R J, Timmis, K N

Two novel conditional broad-host-range cell lysis systems have been developed for the study of natural transformation in bacteria and the environmental fate of DNA released by cell death. Plasmid...

Influence of para-substituents on the oxidative metabolism of o-nitrophenols by Pseudomonas putida B2.

Zeyer, J, Kocher, H P, Timmis, K N

Pseudomonas putida B2 is able to grow on o-nitrophenol (ONP) as the sole source of carbon and nitrogen. ONP was converted by a nitrophenol oxygenase to nitrite and catechol. Catechol was then...

Relatedness of a periplasmic, broad-specificity RNase from Aeromonas hydrophila to RNase I of Escherichia coli and to a family of eukaryotic RNases.

Favre, D, Ngai, P K, Timmis, K N

The isolation, sequencing, and characterization of a periplasmic RNase gene from Aeromonas hydrophila AH1133 is described. Following subcloning of the gene on a 2.7-kb PstI fragment, its direction of...

Three different 2,3-dihydroxybiphenyl-1,2-dioxygenase genes in the gram-positive polychlorobiphenyl-degrading bacterium Rhodococcus globerulus P6.

Asturias, J A, Timmis, K N

Rhodococcus globerulus P6 (previously designated Acinetobacter sp. strain P6, Arthrobacter sp. strain M5, and Corynebacterium sp. strain MB1) is able to degrade a wide range of polychlorinated...

Carbon source-dependent inhibition of xyl operon expression of the Pseudomonas putida TOL plasmid.

Holtel, A, Marqués, S, Möhler, I, Jakubzik, U, Timmis, K N

TOL plasmid-encoded degradation of benzyl alcohol by Pseudomonas putida is inhibited by glucose and other compounds related to the main carbohydrate metabolism in Pseudomonas species. We report here...

Transcriptional induction kinetics from the promoters of the catabolic pathways of TOL plasmid pWW0 of Pseudomonas putida for metabolism of aromatics.

Marqués, S, Holtel, A, Timmis, K N, Ramos, J L

We determined, under several growth conditions, the kinetics of mRNA synthesis from the four Pseudomonas putida pWW0 plasmid promoters involved in degradation of xylenes and methylbenzyl alcohols via...

Genetic evidence that the XylS regulator of the Pseudomonas TOL meta operon controls the Pm promoter through weak DNA-protein interactions.

Kessler, B, Herrero, M, Timmis, K N, De Lorenzo, V

The activation of the Pm promoter of the meta operon of the TOL plasmid of Pseudomonas putida by its cognate XylS activator protein in the presence and absence of benzoate inducers has been examined...

Early and late responses of TOL promoters to pathway inducers: identification of postexponential promoters in Pseudomonas putida with lacZ-tet bicistronic reporters.

De Lorenzo, V, Cases, I, Herrero, M, Timmis, K N

Transcriptional lacZ fusions to the Pu and Pm promoters of the TOL (toluene degradation) plasmid inserted in monocopy in the chromosome of Pseudomonas putida showed a very different responsiveness to...

Characterization of 2,2',3-trihydroxybiphenyl dioxygenase, an extradiol dioxygenase from the dibenzofuran- and dibenzo-p-dioxin-degrading bacterium Sphingomonas sp. strain RW1.

Happe, B, Eltis, L D, Poth, H, Hedderich, R, Timmis, K N

A key enzyme in the degradation pathways of dibenzo-p-dioxin and dibenzofuran, namely, 2,2',3-trihydroxybiphenyl dioxygenase, which is responsible for meta cleavage of the first aromatic ring, has...

Involvement of Pseudomonas putida RpoN sigma factor in regulation of various metabolic functions.

Köhler, T, Harayama, S, Ramos, J L, Timmis, K N

The RpoN protein was originally identified in Escherichia coli as a sigma (sigma) factor essential for the expression of nitrogen regulons. In the present study we cloned the Pseudomonas putida rpoN...

Characterization of five genes in the upper-pathway operon of TOL plasmid pWW0 from Pseudomonas putida and identification of the gene products.

Harayama, S, Rekik, M, Wubbolts, M, Rose, K, Leppik, R A, Timmis, K N

The upper operon of the TOL plasmid pWW0 of Pseudomonas putida encodes a set of enzymes which transform toluene and xylenes to benzoate and toluates. The genetic organization of the operon was...

Regulator and enzyme specificities of the TOL plasmid-encoded upper pathway for degradation of aromatic hydrocarbons and expansion of the substrate range of the pathway.

Abril, M A, Michan, C, Timmis, K N, Ramos, J L

The TOL plasmid upper pathway operon encodes enzymes involved in the catabolism of aromatic hydrocarbons such as toluene and xylenes. The regulator of the gene pathway, the XylR protein, exhibits a...

Roles of the divergent branches of the meta-cleavage pathway in the degradation of benzoate and substituted benzoates.

Harayama, S, Mermod, N, Rekik, M, Lehrbach, P R, Timmis, K N

The TOL plasmid-specified meta-cleavage pathway for the oxidative catabolism of benzoate and toluates branches at the ring cleavage products of catechols and reconverges later at 2-oxopent-4-enoate...

Analysis, cloning, and high-level expression of 2,4-dichlorophenoxyacetate monooxygenase gene tfdA of Alcaligenes eutrophus JMP134.

Streber, W R, Timmis, K N, Zenk, M H

Plasmid pJP4 of Alcaligenes eutrophus JMP134 contains all genes for the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D). Five of these genes, tfdB, tfdC, tfdD, tfdE, and tfdF, have recently...

Vector for regulated expression of cloned genes in a wide range of gram-negative bacteria.

Mermod, N, Ramos, J L, Lehrbach, P R, Timmis, K N

A pKT231-based broad-host-range plasmid vector was constructed which enabled regulation of expression of cloned genes in a wide range of gram-negative bacteria. This vector, pNM185, contained...

Gene order of the TOL catabolic plasmid upper pathway operon and oxidation of both toluene and benzyl alcohol by the xylA product.

Harayama, S, Leppik, R A, Rekik, M, Mermod, N, Lehrbach, P R, Reineke, W, ...

TOL plasmid pWW0 specifies enzymes for the oxidative catabolism of toluene and xylenes. The upper pathway converts the aromatic hydrocarbons to aromatic carboxylic acids via corresponding alcohols...

Mutations leading to constitutive expression from the TOL plasmid meta-cleavage pathway operon are located at the C-terminal end of the positive regulator protein XylS.

Zhou, L M, Timmis, K N, Ramos, J L

The XylS protein is the positive activator of the TOL plasmid meta-cleavage pathway operon for the metabolism of alkylbenzoates in Pseudomonas putida. The regulator stimulates transcription from the...

Characterization of a plasmid-specified pathway for catabolism of isopropylbenzene in Pseudomonas putida RE204.

Eaton, R W, Timmis, K N

A Pseudomonas putida strain designated RE204, able to utilize isopropylbenzene as the sole carbon and energy source, was isolated. Tn5 transposon mutagenesis by means of the suicide transposon donor...

Spontaneous deletion of a 20-kilobase DNA segment carrying genes specifying isopropylbenzene metabolism in Pseudomonas putida RE204.

Eaton, R W, Timmis, K N

The genes encoding isopropylbenzene metabolism in Pseudomonas putida RE204 are readily lost in two ways: by loss (curing) of plasmid pRE4 which specifies the catabolic pathway and by deletion from...

Highly repressible expression system for cloning genes that specify potentially toxic proteins.

O'Connor, C D, Timmis, K N

A highly repressible expression vector system that allows the cloning of potentially deleterious genes has been constructed. Undesired expression of a cloned gene was prevented (i) at the level of...

Transposon mutagenesis analysis of meta-cleavage pathway operon genes of the TOL plasmid of Pseudomonas putida mt-2.

Harayama, S, Lehrbach, P R, Timmis, K N

Hybrid plasmids containing the regulated meta-cleavage pathway operon of TOL plasmid pWWO were mutagenized with transposon Tn1000 or Tn5. The resulting insertion mutant plasmids were examined for...

Transposon mutagenesis and cloning analysis of the pathways for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP134(pJP4).

Don, R H, Weightman, A J, Knackmuss, H J, Timmis, K N

Plasmid pJP4 permits its host bacterium, strain JMP134, to degrade and utilize as sole sources of carbon and energy 3-chlorobenzoate and 2,4-dichlorophenoxyacetic acid (R. H. Don and J. M. Pemberton,...

Enzyme recruitment in vitro: use of cloned genes to extend the range of haloaromatics degraded by Pseudomonas sp. strain B13.

Lehrbach, P R, Zeyer, J, Reineke, W, Knackmuss, H J, Timmis, K N

DNA fragments containing the xylD and xylL genes of TOL plasmid pWW0 -161 of Pseudomonas putida, which code for the catabolic enzymes toluate 1,2-dioxygenase and dihydrodihydroxybenzoic acid...

Replication control mutations of plasmid R6-5 and their effects on interactions of the RNA-I control element with its target.

Brady, G, Frey, J, Danbara, H, Timmis, K N

Nine high copy number mutations of plasmid R6-5, representing five phenotypically distinct groups, have been identified by DNA sequencing. In each mutant plasmid examined, a single nucleotide change...

Localization and functional analysis of transposon mutations in regulatory genes of the TOL catabolic pathway.

Franklin, F C, Lehrbach, P R, Lurz, R, Rueckert, B, Bagdasarian, M, Timmis, K N

Mutant derivatives of the TOL plasmid pWW0-161, containing Tn5 insertions in the xylS and xylR regulatory genes of the catabolic pathway, have been identified and characterized. The two genes are...

Microbial mineralization of ring-substituted anilines through an ortho-cleavage pathway.

Zeyer, J, Wasserfallen, A, Timmis, K N

Moraxella sp. strain G is able to utilize as sole source of carbon and nitrogen aniline, 4-fluoroaniline, 2-chloroaniline, 3-chloroaniline, 4-chloroaniline (PCA), and 4-bromoaniline but not...

Use of cloned genes of Pseudomonas TOL plasmid to effect biotransformation of benzoates to cis-dihydrodiols and catechols by Escherichia coli cells.

Zeyer, J, Lehrbach, P R, Timmis, K N

DNA fragments containing the xylD and xylL genes, which specify the broad-specificity enzymes toluate-1,2-dioxygenase and 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid dehydrogenase, respectively, of...

Construction of stable LamB-Shiga toxin B subunit hybrids: analysis of expression in Salmonella typhimurium aroA strains and stimulation of B subunit-specific mucosal and serum antibody responses.

Su, G F, Brahmbhatt, H N, Wehland, J, Rohde, M, Timmis, K N

The complete Shiga toxin B subunit and two N-terminal segments of the B subunit have been inserted into a cell surface exposed loop of the LamB protein, and expression of the hybrid proteins from...

Fibronectin-binding protein of Streptococcus pyogenes: sequence of the binding domain involved in adherence of streptococci to epithelial cells.

Talay, S R, Valentin-Weigand, P, Jerlström, P G, Timmis, K N, Chhatwal, G S

The sequence of the fibronectin-binding domain of the fibronectin-binding protein of Streptococcus pyogenes (Sfb protein) was determined, and its role in streptococcal adherence was investigated by...

Specific lung mucosal and systemic immune responses after oral immunization of mice with Salmonella typhimurium aroA, Salmonella typhi Ty21a, and invasive Escherichia coli expressing recombinant pertussis toxin S1 subunit.

Walker, M J, Rohde, M, Timmis, K N, Guzmán, C A

Pertussis toxin (PT) is considered an essential protective component for incorporation into new generation vaccines against Bordetella pertussis, the causative agent of whooping cough. Traditionally,...

Production of recombinant Bordetella pertussis serotype 2 fimbriae in Bordetella parapertussis and Bordetella bronchiseptica: utility of Escherichia coli gene expression signals.

Walker, M J, Guzmán, C A, Rohde, M, Timmis, K N

Serotype-specific fimbriae of Bordetella pertussis are considered potential components of new-generation vaccines against whooping cough. Attempts to characterize fimbriae, and indeed other virulence...

Intercellular spread of Shigella flexneri through a monolayer mediated by membranous protrusions and associated with reorganization of the cytoskeletal protein vinculin.

Kadurugamuwa, J L, Rohde, M, Wehland, J, Timmis, K N

The spread of Shigella flexneri in a monolayer of infected Henle and HeLa cells was studied by using immunofluorescence and electron microscopy. Infected cells produced numerous bacterium-containing...

Direct expression of Bordetella pertussis filamentous hemagglutinin in Escherichia coli and Salmonella typhimurium aroA.

Guzmán, C A, Walker, M J, Rohde, M, Timmis, K N

Nonfused (i.e., nonhybrid) filamentous hemagglutinin (FHA) of Bordetella pertussis was efficiently expressed in Escherichia coli K-12 and Salmonella typhimurium aroA at levels higher than those found...

Construction of minitransposons for constitutive and inducible expression of pertussis toxin in bvg-negative Bordetella bronchiseptica.

Walker, M J, Rohde, M, Wehland, J, Timmis, K N

Appropriately detoxified pertussis toxin (PT) of Bordetella pertussis is considered to be an essential component of new-generation whooping cough vaccines, but the development of a procedure to...

Characterization of murine monoclonal antibodies that recognize defined epitopes of pertussis toxin and neutralize its toxic effect on Chinese hamster ovary cells.

Walker, M J, Wehland, J, Timmis, K N, Raupach, B, Schmidt, M A

Three murine monoclonal antibodies (MAb), E19, E205, and E251, raised against pertussis toxin reacted in Western blots (immunoblots) with the S1, S4, and S2-S3 subunits, respectively, and neutralized...

Antibody responses in the lungs of mice following oral immunization with Salmonella typhimurium aroA and invasive Escherichia coli strains expressing the filamentous hemagglutinin of Bordetella pertussis.

Guzmán, C A, Brownlie, R M, Kadurugamuwa, J, Walker, M J, Timmis, K N

The filamentous hemagglutinin (FHA) of Bordetella pertussis was expressed in the attenuated aroA mutant of Salmonella typhimurium, SL3261, and in a strain of Escherichia coli harboring Shigella...

Cloning and expression in Escherichia coli of the Ibc protein genes of group B streptococci: binding of human immunoglobulin A to the beta antigen.

Cleat, P H, Timmis, K N

Total-cell DNA isolated from a highly virulent serotype Ic strain of a group B streptococcus was used to construct a gene bank with bacteriophage lambda L47.1 in Escherichia coli K-12. Recombinant...

Localization of stx, a determinant essential for high-level production of shiga toxin by Shigella dysenteriae serotype 1, near pyrF and generation of stx transposon mutants.

Sekizaki, T, Harayama, S, Brazil, G M, Timmis, K N

Hfr strains of Shigella dysenteriae serotype 1 were constructed by transient integration of an RP4 plasmid derivative carrying transposon Tn501 into the Shigella chromosome through Tn501-mediated...

Small virulence plasmid of Shigella dysenteriae 1 strain W30864 encodes a 41,000-dalton protein involved in formation of specific lipopolysaccharide side chains of serotype 1 isolates.

Watanabe, H, Nakamura, A, Timmis, K N

A 6-megadalton plasmid, pHW400, of Shigella dysenteriae 1 strain W30864 was previously found to specify one or more functions for O-antigen production and bacterial virulence (H. Watanabe and K. N....

Monoclonal antibody detection of IncF group plasmid-encoded TraT protein in clinical isolates of Escherichia coli.

Bitter-Suermann, D, Peters, H, Jürs, M, Nehrbass, R, Montenegro, M, Timmis, K N

The TraT protein specified by IncF group plasmids mediates surface exclusion and bacterial resistance to the lethal activities of serum. In this study, an anti-TraT protein monoclonal antibody was...

Cloning of enterotoxin gene from Aeromonas hydrophila provides conclusive evidence of production of a cytotonic enterotoxin.

Chakraborty, T, Montenegro, M A, Sanyal, S C, Helmuth, R, Bulling, E, Timmis, K N

Culture filtrates of two Aeromonas hydrophila strains which were isolated from patients with diarrhea and assumed to be causative agents of the infections were shown to contain enterotoxic,...

A plasmid-encoded outer membrane protein, TraT, enhances resistance of Escherichia coli to phagocytosis.

Agüero, M E, Aron, L, DeLuca, A G, Timmis, K N, Cabello, F C

The presence of the outer membrane protein TraT, encoded by plasmid R6-5, reduces the sensitivity of Escherichia coli cells to phagocytosis by macrophages. This effect is independent of the bacterial...

Cloning and genetic analysis of serotype 5 M protein determinant of group A streptococci: evidence for multiple copies of the M5 determinant in the Streptococcus pyogenes genome.

Kehoe, M A, Poirier, T P, Beachey, E H, Timmis, K N

A gene bank of group A Streptococcus strain Manfredo (M protein serotype 5) was constructed with a bacteriophage lambda vector-Escherichia coli K-12 host system and screened by immunoblotting hybrid...

Expression of protective and cardiac tissue cross-reactive epitopes of type 5 streptococcal M protein in Escherichia coli.

Poirier, T P, Kehoe, M A, Dale, J B, Timmis, K N, Beachey, E H

The immunochemical properties of type 5 M protein antigens that were expressed in Escherichia coli K-12 by recombinant lambda bacteriophages isolated from a gene bank of serotype 5 Streptococcus...

Monoclonal antibodies to enterobacterial common antigen and to Escherichia coli lipopolysaccharide outer core: demonstration of an antigenic determinant shared by enterobacterial common antigen and E. coli K5 capsular polysaccharide.

Peters, H, Jürs, M, Jann, B, Jann, K, Timmis, K N, Bitter-Suermann, D

We established hybridoma cell lines producing monoclonal antibodies against enterobacterial common antigen (ECA) and a substructure of the outer core of different Escherichia coli lipopolysaccharides...

A small plasmid in Shigella dysenteriae 1 specifies one or more functions essential for O antigen production and bacterial virulence.

Watanabe, H, Timmis, K N

The role of plasmids in the virulence of Shigella dysenteriae 1 W30864, which contains at least five species, was investigated. By means of a standard plasmid-curing procedure, that is, bacterial...

Cloning and expression in Escherichia coli of the streptolysin O determinant from Streptococcus pyogenes: characterization of the cloned streptolysin O determinant and demonstration of the absence of substantial homology with determinants of other thiol-activated toxins.

Kehoe, M, Timmis, K N

A gene bank of Streptococcus pyogenes Richards was constructed in Escherichia coli by using the bacteriophage replacement vector lambda L47.1, and hybrid phage expressing streptolysin O (SLO) were...

Cloning and analysis of the K1 capsule biosynthesis genes of Escherichia coli: lack of homology with Neisseria meningitidis group B DNA sequences.

Echarti, C, Hirschel, B, Boulnois, G J, Varley, J M, Waldvogel, F, Timmis, K N

Genes coding for production of the K1 polysaccharide capsule of Escherichia coli have been cloned. Complementation, insertion, and deletion analyses were used to localize the K1 genes and...

Location of two relaxation nick sites in R6K and single sites in pSC101 and RSF1010 close to origins of vegetative replication: implication for conjugal transfer of plasmid deoxyribonucleic acid.

Nordheim, A, Hashimoto-Gotoh, T, Timmis, K N

A nick-labeling method has been used to localize the relaxation complex nick sites in three plasmids (pSC101, RSF1010, and R6K) that differ markedly in their host range, deoxyribonucleic acid...

Plasmid replication functions: two distinct segments of plasmid R1, RepA and RepD, express incompatibility and are capable of autonomous replication.

Danbara, H, Timmis, J K, Lurz, R, Timmis, K N

The genetic determinants for replication and incompatibility of plasmid R1 were investigated by gene cloning methods, and three types of R1 miniplasmid derivatives were generated. The first,...

Antibody responses in the serum and respiratory tract of mice following oral vaccination with liposomes coated with filamentous hemagglutinin and pertussis toxoid.

Guzman, C A, Molinari, G, Fountain, M W, Rohde, M, Timmis, K N, Walker, M J

Mice were orally vaccinated with liposomes coated with filamentous hemagglutinin (FHA) and detoxified pertussis toxin (PT) of Bordetella pertussis. FHA- and PT-specific immunoglobulin G (IgG) was...

Invasion and intracellular survival of Bordetella bronchiseptica in mouse dendritic cells.

Guzman, C A, Rohde, M, Bock, M, Timmis, K N

We have studied the interaction between the respiratory pathogen Bordetella bronchiseptica and murine spleen dendritic cells, important antigen-presenting cells that are found in the airway...

Mechanisms involved in uptake of Bordetella bronchiseptica by mouse dendritic cells.

Guzman, C A, Rohde, M, Timmis, K N

The invasion and intracellular survival of Bordetella bronchiseptica in mouse dendritic cells were investigated. The results obtained suggest that B. bronchiseptica binds specifically to glycosylated...

Metagenome Survey of Biofilms in Drinking-Water Networks

Schmeisser, C., Stöckigt, C., Raasch, C., Wingender, J., Timmis, K. N., Wenderoth, D. F., ...

Most naturally occurring biofilms contain a vast majority of microorganisms which have not yet been cultured, and therefore we have little information on the genetic information content of these...

Upstream binding sequences of the XylR activator protein and integration host factor in the xylS gene promoter region of the Pseudomonas TOL plasmid.

Holtel, A, Timmis, K N, Ramos, J L

The xylR and xylS genes, which encode the positive regulators of the TOL plasmid catabolic pathways, are adjacent genes on the TOL plasmid and are transcribed from divergent promoters. Transcription...

Bacterial infection of wounds: fibronectin-mediated adherence group A and C streptococci to fibrin thrombi in vitro.

Chhatwal, G S, Valentin-Weigand, P, Timmis, K N

Adherence of group A, B, and C streptococci to fibrin thrombi was studied by using a novel fluorochrome microassay carried out in microdilution plates in which fibrin thrombi had been prepared by...

Regulation of DNA replication: "target" determinant of the replication control elements of plasmid R6-5 lies within a control element gene.

Danbara, H, Brady, G, Timmis, J K, Timmis, K N

The replication control system of plasmid R6-5 has been investigated by characterization of high-copy-number mutant miniplasmids, development of an in vivo assay for the site of action or "target" of...

A family of positive regulators related to the Pseudomonas putida TOL plasmid XylS and the Escherichia coli AraC activators.

Ramos, J L, Rojo, F, Zhou, L, Timmis, K N

The XylS family consists of a least 8 different transcriptional regulators. Six of these proteins are positive regulators for the catabolism of carbon sources (benzoate and sugars) in Escherichia...

Cloning of chicken lysozyme structural gene sequences synthesized in vitro.

Sippel, A E, Land, H, Lindenmaier, W, Nguyen-Huu, M C, Wurtz, T, Timmis, K N, ...

Double-stranded chicken lysozyme cDNA was synthesized from an oviduct mRNA fraction enriched for lysozyme mRNA. The ds-cDNA was inserted into the BamHI site of plasmid pBR322 using chemically...

Plasmid replication functions. IV. Promoters in the replication region of plasmid R6-5.

Slocombe, P M, Ely, S, Timmis, K N

Eight RNA polymerase binding sites have been shown to map within the EcoRI fragment E-2 (replication region RepA EcoRI fragment) of plasmid R6-5 and all but one have been shown to contain active...

Molecular and functional analysis of the TOL plasmid pWWO from Pseudomonas putida and cloning of genes for the entire regulated aromatic ring meta cleavage pathway.

Franklin, F C, Bagdasarian, M, Bagdasarian, M M, Timmis, K N

The genetic organization of the Pseudomonas putida plasmid pWWO-161, which encodes enzymes for the degradation of toluene and related aromatic hydrocarbons, has been investigated by transposition...

An inhibitor of SOS induction, specified by a plasmid locus in Escherichia coli.

Bagdasarian, M, Bailone, A, Bagdasarian, M M, Manning, P A, Lurz, R, Timmis, K N, ...

Plasmid R6-5 contains a locus whose product inhibits induction of sfiA and prophage lambda in a recA441 mutant at 42 degrees C and in a recA+ host after treatment with nalidixic acid. This plasmidic...

Altered effector specificities in regulators of gene expression: TOL plasmid xylS mutants and their use to engineer expansion of the range of aromatics degraded by bacteria.

Ramos, J L, Stolz, A, Reineke, W, Timmis, K N

Stimulation of transcription from positively regulated promoters involves regulatory proteins that have been activated, generally as a consequence of binding low molecular weight effector molecules....

NZB mouse system for production of monoclonal antibodies to weak bacterial antigens: isolation of an IgG antibody to the polysaccharide capsules of Escherichia coli K1 and group B meningococci.

Frosch, M, Görgen, I, Boulnois, G J, Timmis, K N, Bitter-Suermann, D

A system for the production of monoclonal antibodies, particularly of the IgG type, against weakly immunogenic bacterial polysaccharide antigens is described. This system, which is based on the...

An upstream XylR- and IHF-induced nucleoprotein complex regulates the sigma 54-dependent Pu promoter of TOL plasmid.

De Lorenzo, V, Herrero, M, Metzke, M, Timmis, K N

Transcription from promoter Pu of the upper catabolic operon of the Pseudomonas putida TOL plasmid which specifies conversion of toluene/xylenes to benzoate/toluates is activated by the TOL-encoded...

Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria.

Herrero, M, De Lorenzo, V, Timmis, K N

A simple procedure for cloning and stable insertion of foreign genes into the chromosomes of gram-negative eubacteria was developed by combining in two sets of plasmids (i) the transposition features...

Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria.

De Lorenzo, V, Herrero, M, Jakubzik, U, Timmis, K N

A collection of Tn5-derived minitransposons has been constructed that simplifies substantially the generation of insertion mutants, in vivo fusions with reporter genes, and the introduction of...

Plasmid-determined resistance to serum bactericidal activity: a major outer membrane protein, the traT gene product, is responsible for plasmid-specified serum resistance in Escherichia coli.

Moll, A, Manning, P A, Timmis, K N

Resistance to the bactericidal activity of serum appears to be an important virulence property of invasive bacteria. The conjugative multiple-antibiotic-resistance plasmid R6-5 was found to confer...

Transcription of the TOL plasmid toluate catabolic pathway operon of Pseudomonas putida is determined by a pair of co-ordinately and positively regulated overlapping promoters.

Mermod, N, Lehrbach, P R, Reineke, W, Timmis, K N

Expression of the meta-cleavage pathway operon of TOL plasmid pWW0 of Pseudomonas putida is positively regulated by the xylS gene product. We have sequenced the promoter region of this operon and...

Tracking the Response of Burkholderia cepacia G4 5223-PR1 in Aquifer Microcosms

Winkler, J., Timmis, K. N., Snyder, R. A.

The introduction of bacteria into the environment for bioremediation purposes (bioaugmentation) requires analysis and monitoring of microbial population dynamics to define persistence and activity...

Formation of Dimethylmuconolactones from Dimethylphenols by Alcaligenes eutrophus JMP 134

Pieper, D. H., Stadler-Fritzsche, K., Knackmuss, H., Timmis, K. N.

2,3-, 2,4-, 2,5-, 3,4-, and 3,5-dimethylphenols were cometabolized by 2,4-dichlorophenoxyacetate-grown Alcaligenes eutrophus JMP 134 or the constitutive derivative JMP 134-1 via the ortho pathway...

Degradation of Chlorinated Dibenzofurans and Dibenzo-p-Dioxins by Sphingomonas sp. Strain RW1

Wilkes, H., Wittich, R., Timmis, K. N., Fortnagel, P., Francke, W.

The ability of the dibenzofuran- and dibenzo-p-dioxin-mineralizing bacterium Sphingomonas sp. strain RW1 (R.-M. Wittich, H. Wilkes, V. Sinnwell, W. Francke, and P. Fortnagel, Appl. Environ....

Use of Colicin E3 for Biological Containment of Microorganisms

Munthali, M. T., Timmis, K. N., Diaz, E.

The genetic determinant of the lethal antibiotic colicin E3 was cloned under the control of a tightly regulated promoter in the absence of the gene for its cognate inhibitor. Combination of this...

Metabolism of Chlorotoluenes by Burkholderia sp. Strain PS12 and Toluene Dioxygenase of Pseudomonas putida F1: Evidence for Monooxygenation by Toluene and Chlorobenzene Dioxygenases

Lehning, A., Fock, U., Wittich, R., Timmis, K. N., Pieper, D. H.

The degradation of toluene by Pseudomonas putida F1 and of chlorobenzenes by Burkholderia sp. strain PS12 is initiated by incorporation of dioxygen into the aromatic nucleus to form...

Mineralization of 4-Chlorodibenzofuran by a Consortium Consisting of Sphingomonas sp. Strain RW1 and Burkholderia sp. Strain JWS

Arfmann, H., Timmis, K. N., Wittich, R.

The dibenzofuran-degrading bacterium Sphingomonas sp. strain RW1 (R.-M. Wittich, H. Wilkes, V. Sinnwell, W. Francke, and P. Fortnagel, Appl. Environ. Microbiol. 58:1005-1010, 1992) attacks...

Evidence that Formation of Protoanemonin from Metabolites of 4-Chlorobiphenyl Degradation Negatively Affects the Survival of 4-Chlorobiphenyl-Cometabolizing Microorganisms

Blasco, R., Mallavarapu, M., Wittich, R., Timmis, K. N., Pieper, D. H.

A rapid decline in cell viability of different PCB-metabolizing organisms was observed in soil microcosms amended with 4-chlorobiphenyl. The toxic effect could not be attributed to 4-chlorobiphenyl...

Reduction in diversity of the colonic mucosa associated bacterial microflora in patients with active inflammatory bowel disease

Ott, S J, Musfeldt, M, Wenderoth, D F, Hampe, J, Brant, O, Fölsch, U R, ...

Background and aims: The intestinal bacterial microflora plays an important role in the aetiology of inflammatory bowel disease (IBD). As most of the colonic bacteria cannot be identified by culture...

Shift from Acetoclastic to H2-Dependent Methanogenesis in a West Siberian Peat Bog at Low pH Values and Isolation of an Acidophilic Methanobacterium Strain▿

Kotsyurbenko, O. R., Friedrich, M. W., Simankova, M. V., Nozhevnikova, A. N., Golyshin, P. N., Timmis, K. N., ...

Methane production and archaeal community composition were studied in samples from an acidic peat bog incubated at different temperatures and pH values. H2-dependent methanogenesis increased strongly...

Crystal structure of cis-biphenyl-2,3-dihydrodiol-2,3-dehydrogenase from a PCB degrader at 2.0 A resolution.

Hülsmeyer, M., Hecht, H. J., Niefind, K., Hofer, B., Eltis, L. D., Timmis, K. N., ...

cis-Biphenyl-2,3-dihydrodiol-2,3-dehydrogenase (BphB) is involved in the aerobic biodegradation of polychlorinated biphenyls (PCBs). The crystal structure of the NAD+-enzyme complex was determined by...