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nucleotide homology and organization of chlorocatechol oxidation genes of plasmids pjp4 and pac27.the 2,4-dichlorophenoxyacetate (2,4-d) catabolic plasmid pjp4 of alcaligenes eutrophus jmp134 contains two sets of nonidentical chlorocatechol oxidation gene sequences physically separated by a 7 kb dna region. we determined the nucleotide sequence of the 1.6 kb hindiii fragment containing the known genes tfdc and tfdd (don et al. 1985) which encode pyrocatechase and cycloisomerase, respectively. the 1.3 kb bglii-hindiii segment of recombinant plasmid pdc25 containing at least three chlorocatech ...19882830460
cloning, dna sequence analysis, and expression in escherichia coli of the gene for mandelate racemase from pseudomonas putida.the gene for mandelate racemase (ec 5.1.2.2) from pseudomonas putida (atcc 12633) was cloned in pseudomonas aeruginosa (atcc 15692). the selection for the cloned gene was based upon the inability of p. aeruginosa to grow on (r)-mandelate as sole carbon source by virtue of the absence of mandelate racemase in its mandelate pathway. fragments of p. putida dna obtained by digestion of chromosomal dna with sau3a were ligated into the bamhi site of the gram-negative vector pkt230 and transformed into ...19882831968
variation in the ability of pseudomonas sp. strain b13 cultures to utilize meta-chlorobenzoate is associated with tandem amplification and deamplification of dna.single-colony isolates of pseudomonas sp. strain b13 were examined for their ability to utilize benzoate (ben) and meta-chlorobenzoate (3cb) as the sole carbon source. scoring of b13 cultures by the replica-plating technique revealed that under nonselective conditions, b13 spontaneously formed four different types of colonies: 3cb+ ben+, 3cb+ ben-, 3cb- ben-, 3cb- ben+. successive testing of each of the four colony types showed that each produced the same four different types of single-colony is ...19882832387
variation in the ability of pseudomonas sp. strain b13 cultures to utilize meta-chlorobenzoate is associated with tandem amplification and deamplification of dna.single-colony isolates of pseudomonas sp. strain b13 were examined for their ability to utilize benzoate (ben) and meta-chlorobenzoate (3cb) as the sole carbon source. scoring of b13 cultures by the replica-plating technique revealed that under nonselective conditions, b13 spontaneously formed four different types of colonies: 3cb+ ben+, 3cb+ ben-, 3cb- ben-, 3cb- ben+. successive testing of each of the four colony types showed that each produced the same four different types of single-colony is ...19882832387
enumeration of tn5 mutant bacteria in soil by using a most- probable-number-dna hybridization procedure and antibiotic resistance.investigations were made into the utility of dna hybridization in conjunction with a microdilution most-probable-number procedure for the enumeration of rhizobium spp. and pseudomonas putida in soil. isolates of rhizobium spp. and p. putida carrying the transposon tn5 were added to sterile and nonsterile burbank sandy loam soil and enumerated over time. soil populations of rhizobia were enumerated by colony hybridization, most-probable-number-dna hybridization procedure, plate counts, plant infe ...19882833161
dehalogenase genes of pseudomonas putida pp3 on chromosomally located transposable elements.pseudomonas putida pp3 utilizes halogenated alkanoic acids (haa) such as 2,2-dcpa as its sole carbon and energy sources. spontaneous hha- mutants, isolated by selection for resistance to the toxic analogs monochloroacetic acid and dichloroacetic acid, arose at frequencies several orders of magnitude higher than expected for spontaneous mutations. analysis of the five classes of mutants isolated suggested that the dehalogenase and haa permease genes were on chromosomally located transposable elem ...19852835577
cosmid cloning of five zymomonas trp genes by complementation of escherichia coli and pseudomonas putida trp mutants.a library of zymomonas mobilis genomic dna was constructed in the broad-host-range cosmid plafr1. the library was mobilized into a variety of escherichia coli and pseudomonas putida trp mutants by using the helper plasmid prk2013. five z. mobilis trp genes were identified by the ability to complement the trp mutants. the trpf, trpb, and trpa genes were on one cosmid, while the trpd and trpc genes were on two separate cosmids. the organization of the z. mobilis trp genes seems to be similar to th ...19882838460
benzoate-dependent induction from the op2 operator-promoter region of the tol plasmid pwwo in the absence of known plasmid regulatory genes.expression of the lower catabolic pathway of the tol plasmid pwwo requires an aromatic acid inducer and the product of the xyls regulatory gene. pseudomonas putida cells transformed with a plasmid containing the operator-promoter region of the lower pathway (op2 [or pm]), upstream from the catechol 2,3-dioxygenase structural gene, showed enzyme induction in the absence of known tol plasmid regulatory genes. induction was not seen in transformed escherichia coli cells or in a p. putida mutant lac ...19882841300
klebsiella pneumoniae origin of replication (oric) is not active in caulobacter crescentus, pseudomonas putida, and rhodobacter sphaeroides.a dna fragment carrying genes encoding the conjugal transfer system of the broad host range plasmid rk2 was inserted into a plasmid carrying the chromosomal origin of replication (oric) from klebsiella pneumoniae. the resulting plasmid, peon1, was readily transferred between gram-negative bacteria and carried two potential origins of replication: oric and the replication origin from pbr322 (oripbr). although peon1 could be transferred to caulobacter crescentus, pseudomonas putida, and rhodobacte ...19882841304
isolation and characterization of a new plasmid from a flavobacterium sp. which carries the genes for degradation of 2,4-dichlorophenoxyacetate.a flavobacterium sp. (strain 50001), capable of degrading 2,4-dichlorophenoxyacetate (2,4-d), 2-methyl-4-chlorophenoxyacetate, and 2-chlorobenzoate and imparting resistance to mercury, harbored a degradative plasmid, prc10. cured strains of the flavobacterium sp. lost the plasmid as well as the ability to degrade these chlorinated compounds. comparison of this plasmid with the well-characterized 2,4-d-degradative plasmid pjp4 from alcaligenes eutrophus showed regions of homology between the two ...19882842290
organization and multiple regulation of histidine utilization genes in pseudomonas putida.the arrangement of the histidine utilization (hut) genes in pseudomonas putida was established by examining the structure of a dna segment that had been cloned into escherichia coli via a cosmid vector. southern blot analysis revealed that the restriction patterns of the hut genes cloned into e. coli and present in the p. putida genome were identical, indicating that no detectable dna rearrangement took place during the cloning. expression of the hut genes from a series of overlapping clones ind ...19882842309
toluene degradation by pseudomonas putida f1: genetic organization of the tod operon.pseudomonas putida ppf1 degrades toluene through cis-toluene dihydrodiol to 3-methylcatechol. the latter compound is metabolized through the well-established meta pathway for catechol degradation. the first four steps in the pathway involve the sequential action of toluene dioxygenase (todabc1c2), cis-toluene dihydrodiol dehydrogenase (todd), 3-methylcatechol 2,3-dioxygenase (tode), and 2-hydroxy-6-oxo-2,4-heptadienoate hydrolase (todf). the genes for these enzymes form part of the tod operon wh ...19882843094
[interaction of heterologous bacteriophages: growth suppression of temperate pp56 phage of pseudomonas putida by the transposable phage d3112 of pseudomonas aeruginosa].we have found an inhibiting effect of hybrid rp4::d3112 plasmid (where d3112 is represented as genome of a transposable phage specific for pseudomonas aeruginosa) on the development of temperate p. putida phage pp56. the study of the effect has revealed a previously unknown locus (in the region 12-14.2 kb of the d3112 genome) which functions in the prophage state. the locus affects pp56 decreasing phage yield. mutants of pp56 insensitive to inhibition were found.19882843421
alkane utilization in pseudomonas oleovorans. structure and function of the regulatory locus alkr.the oct plasmid-localized alkbac operon encodes enzymes for alkane hydroxylation and alkanol dehydrogenation. the positively controlled expression of the operon is very efficient in both pseudomonas putida and escherichia coli. two regulatory functions have been ascribed to the regulatory locus alkr: inducer recognition and transcriptional activation of the operon. we have cloned and localized the alkr locus on a 4.9-kilobase pair sali fragment. the alkr region was analyzed for translation produ ...19882843518
construction of an expression vector for the fission yeast schizosaccharomyces pombe.we have isolated and characterized a s. pombe promoter using a functional heterologous gene product assay. random s. pombe genomic fragments were cloned upstream from the promoterless 'lacz gene and tested in vivo for their efficiency to promote expression of the beta-galactosidase protein in the fission yeast. an efficient s. pombe promoter called 54/1 was isolated and shown to drive up to 5% of total protein synthesis as beta-galactosidase. the structure and nucleotide sequence of this promote ...19882843820
[cloning and localization of the replication and mobilization regions in the d-plasmid of pseudomonas putida pbs286 (incp-9) with a broad host range].rep-mob loci of naphthalene degradative plasmid pbs286 (incp-9) have been cloned on the escherichia coli vectors puc19 and pubr322. these loci confer to recombinant plasmids pbs952 and pbs953 the ability for effective mobilization by rp4 (incp-1) and f plasmid, as well as constant maintenance in various gram-negative bacteria. localization of cloned sequences in the restriction fragments of conservative part of the pbs286 genome was established. the data obtained correlate with the analysis of p ...19882844624
cloning and expression in escherichia coli of histidine utilization genes from pseudomonas putida.a library of the pseudomonas putida chromosome, prepared through the use of the cosmid pjb8 ligated to a partial sau3a digest of bacterial dna, followed by in vitro packaging into bacteriophage lambda particles, was used to construct a strain of escherichia coli which contained the genes for histidine utilization. this isolate produced a repressor product and all five enzymes required in pseudomonas spp. for histidine dissimilation, whereas none of these could be detected in the nontransduced pa ...19852858467
in vivo role of sulfite in photocontrol of urocanase from pseudomonas putida. 19852858892
activation of urocanase from pseudomonas putida by electronically excited triplet species.urocanase from pseudomonas putida becomes inactive in growing and resting cells and, as shown previously, is activated by the direct absorption of ultraviolet light. in this study, we describe the activation of urocanase by energy transfer from triplet indole-3-aldehyde, generated in the peroxidase-catalyzed aerobic oxidation of indole-3-acetic acid. the activation was time-, temperature-, and ph-dependent. the involvement of reactive oxygen intermediates was excluded by the lack of effect of ap ...19852864338
tryptophanyl fluorescence quenching of urocanase from pseudomonas putida by acrylamide, cesium, iodide, and imidazolepropionate. 19852864711
effect of temperature on sulfite-mediated dark reversion of urocanase in pseudomonas putida. 19872888143
phenoxyacetic acid degradation by the 2,4-dichlorophenoxyacetic acid (tfd) pathway of plasmid pjp4: mapping and characterization of the tfd regulatory gene, tfdr.plasmid pjp4 enables alcaligenes eutrophus jmp134 to degrade 3-chlorobenzoate and 2,4-dichlorophenoxyacetic acid (tfd). plasmid pro101 is a derivative of pjp4 obtained by insertion of tn1721 into a nonessential region of pjp4. plasmid pro101 was transferred by conjugation to several pseudomonas strains and to a. eutrophus aeo106, a cured isolate of jmp134. aeo106(pro101) and some pseudomonas transconjugants grew on tfd. transconjugants with a chromosomally encoded phenol hydroxylase also degrade ...19892914848
isolation of a third lipoamide dehydrogenase from pseudomonas putida.pseudomonads are the only organisms so far known to produce two lipoamide dehydrogenases (lpds), lpd-val and lpd-glc. lpd-val is the specific e3 component of branched-chain oxoacid dehydrogenase, and lpd-glc is the e3 component of 2-ketoglutarate and possibly pyruvate dehydrogenases and the l-factor of the glycine oxidation system. three mutants of pseudomonas putida, js348, js350, and js351, affected in lpdg, the gene encoding lpd-glc, have been isolated; all lacked 2-ketoglutarate dehydrogenas ...19892914869
demonstration, characterization, and mutational analysis of nahr protein binding to nah and sal promoters.the nahr gene of plasmid nah7 of pseudomonas putida encodes a 36-kilodalton polypeptide which activates transcription of the nah and sal operons in response to the inducer salicylate. a gel mobility shift assay was used to identify a dna-binding activity which was present only in extracts from either p. putida or escherichia coli containing a functional nahr gene. the binding activity was highly specific for dna containing the nah or sal promoters, but the apparent affinity for the promoters was ...19892914873
effect of degradative plasmid cam-oct on responses of pseudomonas bacteria to uv light.the effect of plasmid cam-oct on responses to uv irradiation was compared in pseudomonas aeruginosa, in pseudomonas putida, and in pseudomonas putida mutants carrying mutations in uv response genes. cam-oct substantially increased both survival and mutagenesis in the two species. p. aeruginosa strains without cam-oct exhibited much higher uv sensitivity than did p. putida strains. uv-induced mutagenesis of plasmid-free p. putida was easily detected in three different assays (two reversion assays ...19892914880
molecular cloning of salicylate hydroxylase genes from pseudomonas cepacia and pseudomonas putida.the sal gene encoding pseudomonas cepacia salicylate hydroxylase was cloned and the sal encoding pseudomonas putida salicylate hydroxylase was subcloned into plasmid vector pro2317 to generate recombinant plasmids ptk3 and ptk1, respectively. both cloned genes were expressed in the host pseudomonas aeruginosa pao1. the parental strain can utilize catechol, a product of the salicylate hydroxylase-catalyzed reaction, but not salicylate as the sole carbon source for growth due to a natural deficien ...19892916843
sequence analysis of the lpdv gene for lipoamide dehydrogenase of branched-chain-oxoacid dehydrogenase of pseudomonas putida.the production of two lipoamide dehydrogenases by pseudomonas is so far unique. one, lpd-val, is the specific e3 component of the branched-chain-oxoacid dehydrogenase and the second, lpd-glc, is the e3 component of 2-oxoglutarate dehydrogenase and the l-factor of the glycine oxidation system. the objective of the present research was to determine the nucleotide sequence of the structural gene for lpd-val in order to compare its deduced amino acid structure with that of other redox-active disulfi ...19892917566
crystal structure of muconolactone isomerase at 3.3 a resolution.the crystal structure of muconolactone isomerase from pseudomonas putida, a unique molecule with ten 96 amino acid subunits and 5-fold, and 2-fold symmetries, has been solved at 3.3 a resolution. the non-crystallographic symmetries were used to refine the initial single isomorphous replacement phases and produce an interpretable 10-fold averaged map. the backbone trace is complete and confirmed by the amino acid sequence fit. each subunit is composed of a body with two alpha-helices and an antip ...19892926818
siderophore-mediated uptake of fe3+ by the plant growth-stimulating pseudomonas putida strain wcs358 and by other rhizosphere microorganisms.under iron-limited conditions, pseudomonas putida wcs358 produces a siderophore, pseudobactin 358, which is essential for the plant growth-stimulating ability of this strain. cells of strain wcs358, provided that they have been grown under fe3+ limitation, take up 55fe3+ from the 55fe3+-labeled pseudobactin 358 complex with km and vmax values of 0.23 microm and 0.14 nmol/mg of cell dry weight per min, respectively. uptake experiments with cells treated with various metabolic inhibitors showed th ...19882971647
cloning of the gene for the common pathogenic neisseria h.8 antigen from neisseria gonorrhoeae.neisseria gonorrhoeae dna that encodes the pathogenic neisseria h.8 common antigen was cloned in the lambda phage sep6. the recombinant phage, designated s6h.8, was detected immunologically with a monoclonal antibody that binds to the h.8 antigen. the gonococcal and s6h.8 forms of the antigen yielded identical partial proteolysis epitope maps. neisseria species that did not manifest the h.8 antigen showed little or no dna homology with s6h.8. this clone should facilitate investigation into the c ...19852981198
purification and properties of ferredoxintol. a component of toluene dioxygenase from pseudomonas putida f1.toluene dioxygenase oxidizes toluene to (+)-cis-1(s),2(r)-dihydroxy-3-methylcyclohexa-3,5-diene. this reaction is catalyzed by a multienzyme system that is induced in cells of pseudomonas putida f1 during growth on toluene. one of the components of toluene dioxygenase has been purified to homogeneity and shown to be an iron-sulfur protein that has been designated ferredoxintol. the molecular weight of ferredoxintol was calculated to be 15,300, and the purified protein was shown to contain 2 g of ...19852982815
sal-tol in vivo recombinant plasmid pkf439.sal-tol in vivo recombinant plasmid pkf439 was characterized in a strain from a mixed culture of bacteria harboring various degradative plasmids. analysis of the gene organization of pkf439 revealed that the 57-kilobase tol fragment, including the 40-kilobase tol metabolic region, was inserted into the complete sal replicon at the position of smai-c within xhoi-b of sal. the molecular size of pkf439 was calculated to be 138 kilobases. pkf439 could be transferred to pseudomonas putida and pseudom ...19852987190
shuttle vector for escherichia coli, pseudomonas putida, and pseudomonas aeruginosa.a hybrid plasmid capable of replication in 2 different genera, escherichia and pseudomonas, was constructed. this plasmid dna can be used as a cloning vector in e. coli and pseudomonades cells. the described hybrid plasmid pld411 has been constructed on the basis of 2 small e. coli vector r-plasmids used in our laboratory and cryptic plasmid pww2 or p. putida mt1. plasmid pld411 dna was mapped with restrictases; its biological activity in transformations of different bacterial strains was studie ...19852990432
p-cresol methylhydroxylase. assay and general properties.p-cresol methylhydroxylase from pseudomonas putida, an anaerobic dehydrogenase that catalyses the oxidation of p-cresol to p-hydroxybenzyl alcohol and then to p-hydroxybenzaldehyde, is an enzyme of great interest in several respects. one of these is the fact that its flavoprotein and cytochrome c subunits may be reversibly dissociated with ease, with full regeneration of the activity and its native properties on recombining the components. bisubstrate kinetic analysis of the unresolved enzyme gi ...19852990444
the site of cyclic amp-dependent protein kinase catalyzed phosphorylation of cytochrome p-450 lm2.the phenobarbital-inducible form of cytochrome p-450 purified from rabbit liver microsomes is phosphorylated by camp-dependent protein kinase at a single site, the serine residue in position 128 of the amino acid sequence. the serine is located in a characteristic recognition sequence for camp-dependent protein kinase and is part of a primary structure which is conserved during evolution, present also in phenobarbital-inducible rat cytochrome and cytochrome p-450 cam from pseudomonas putida. the ...19852991008
p450cam gene cloning and expression in pseudomonas putida and escherichia coli.the gene camc, which encodes the cytochrome p450 monoxygenase protein, was cloned into the shuttle vector pkt240 and recovered as the recombinant pkg201 with a 2.3 kb insert from the cam plasmid in the psti site. the gene product is expressed constitutively in p. putida and in e. coli whereas the inverted insert clone lacks expression, indicating absence of an insert promoter.19852992468
determination of the transcription initiation site and identification of the protein product of the regulatory gene xylr for xyl operons on the tol plasmid.the xylr gene is a regulatory gene on the tol plasmid, which acts in a positive manner on xyl operons for degradation of toluene and xylenes in pseudomonas putida. a dna fragment containing the xylr promoter region was cloned on promoter-probing vectors, and its nucleotide sequence was determined. the transcription initiation site of the xylr gene was determined in cells of p. putida and escherichia coli by s1 nuclease and reverse transcriptase mapping. two initiation sites were detected which w ...19852993247
transposon donor plasmids, based on colib-p9, for use in pseudomonas putida and a variety of other gram negative bacteria.the properties of plg221, a derivative of the colib plasmid carrying the transposon tn5 are described. this plasmid can be used to introduce tn5 by conjugation from escherichia coli into a variety of gram negative bacteria outside the host range for maintenance of colib. plasmid plg221, and a similar plasmid plg223 carrying tn10 may be of general utility as vectors for transposon-mediated mutagenesis in a variety of gram negative bacteria.19852993815
isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating pseudomonas putida wcs358.the plant-growth-stimulating pseudomonas putida wcs358 was mutagenized with transposon tn5. the resulting mutant colony bank was screened for mutants defective in the biosynthesis of the fluorescent siderophore. a total of 28 mutants, divided into six different classes, were isolated that were nonfluorescent or defective in iron acquisition or both. these different types of mutants together with the probable overall structure of the siderophore, i.e., a small peptide chain attached to a fluoresc ...19852997118
evolutionary conservation of genes coding for meta pathway enzymes within tol plasmids pww0 and pww53.pseudomonas putida mt53 contains a tol plasmid, pww53, that encodes toluene-xylene catabolism. pww53 is nonconjugative, is about 105 to 110 kilobase pairs (kbp) in size, and differs significantly in its restriction endonuclease digestion pattern and incompatibility group from the archetypal tol plasmid pww0. an rp4::pww53 cointegrate plasmid, pww53-4, containing about 35 kbp of pww53 dna, including the entire catabolic pathway genes, was formed, and a restriction map for kpni, hindiii, and bamhi ...19852997136
[17o]water and nitric oxide binding by protocatechuate 4,5-dioxygenase and catechol 2,3-dioxygenase. evidence for binding of exogenous ligands to the active site fe2+ of extradiol dioxygenases.pseudomonas testosteroni protocatechuate 4,5-dioxygenase and pseudomonas putida catechol 2,3-dioxygenase (metapyrocatechase) catalyze extradiol-type oxygenolytic cleavage of the aromatic ring of their substrates. the essential active site fe2+ of each enzyme binds nitric oxide (no) to produce an epr active complex with an electronic spin of s = 3/2. hyperfine broadening of the epr resonances of the nitrosyl complexes by 17o-enriched h2o shows that water is bound directly to the fe2+ in the nativ ...19852997190
[expression of pseudomonas aeruginosa transposable phages in pseudomonas putida cells. i. the establishment of a lysogenic state and the effectiveness of lytic development].expression of transposable phages (tp) of pseudomonas aeruginosa in the cells of p. putida was studied. the high efficiency of phage lytic development was shown both as a consequence of zygotic induction after transfer of the rp4::tpc+ plasmid into nonlysogenic recipients, and as a result of heat induction of lysogens ppg1 (d3112cts15). the high phage yield (20-25 particles of d3112cts phage per one cell of p. putida) is an evidence for a high level of transposition in the cells of this bacteria ...19852998927
omega mutagenesis in gram-negative bacteria: a selectable interposon which is strongly polar in a wide range of bacterial species.we have used the 2.0-kb dna fragment omega [prentki and krisch, gene 29 (1984) 303-313] to mutagenize in vitro a broad-host-range plasmid carrying the entire meta-cleavage pathway of the pseudomonas putida tol plasmid pww0. the mutant plasmids were subsequently introduced by conjugal mobilization into a variety of gram-negative bacteria. the omega fragment carries a selectable marker (aada+; spcr/smr), which is expressed in all species tested, as well as flanking transcription and translation te ...19852998930
homology between nucleotide sequences of promoter regions of nah and sal operons of nah7 plasmid of pseudomonas putida.the in vivo transcription start sites of the nah and sal operons of the nah7 plasmid were determined by s1 nuclease mapping and the nucleotide sequence surrounding these transcription start sites was determined. since expression of both of these operons is coordinately controlled by the product of the transcriptional activator gene nahr, the sequences were compared to locate potential sites involved in common regulation. in the 100-base-pair region preceding transcription start sites of both ope ...19863001734
[expression of pseudomonas aeruginosa phage transposons in pseudomonas putida ppg1 cells. ii. zygotic induction--a necessary condition for the formation of defective lysogens].the transfer of hybrid plasmid rp4::pt (where pt is the genome of a transposable phage specific for pseudomonas aeruginosa) into recipient cells of p. putida strain ppg1 occurs with the same frequency as into p. aeruginosa, the homologous host for pt. approximately 1/3 of all ppg1 exconjugants carrying rp4 markers lost the capability to produce viable pt phage. in contrast, in a cross with homologous recipient p. aeruginosa all exconjugant clones contained nondefective prophages in the hybrid pl ...19853002910
cloning and expression of acinetobacter calcoaceticus catbcde genes in pseudomonas putida and escherichia coli.this report describes the isolation and preliminary characterization of a 5.0-kilobase-pair (kbp) ecori dna restriction fragment carrying the catbcde genes from acinetobacter calcoaceticus. the respective genes encode enzymes that catalyze four consecutive reactions in the catechol branch of the beta-ketoadipate pathway: catb, muconate lactonizing enzyme (ec 5.5.1.1); catc, muconolactone isomerase (ec 5.3.3.4); catd, beta-ketoadipate enol-lactone hydrolase (ec 3.1.1.24); and cate, beta-ketoadipa ...19863003031
characterization of the oct plasmid encoding alkane oxidation and mercury resistance in pseudomonas putida.transformation of pseudomonas putida and analysis for plasmid dna revealed that both n-alkane oxidation and mercury resistance are encoded on a single 220-megadalton oct plasmid molecule. derivatives of oct having lost the mercury resistance function could be readily isolated and contained a smaller plasmid estimated to be 170 megadaltons. the results show that segregation of the mercury resistance property occurs not by loss of a separate mer plasmid as previously thought but by a deletion in t ...19863003035
nucleotide sequence of the pseudomonas putida cytochrome p-450cam gene and its expression in escherichia coli.cytochrome p-450cam catalyzes the stereospecific methylene hydroxylation of camphor to form 5-exohydroxycamphor and is encoded by the camc gene on the cam plasmid of pseudomonas putida, atcc 17453. the cytochrome p-450cam structural gene has been cloned by mutant complementation in p. putida (koga, h., rauchfuss, b., and gunsalus, i. c. (1985) biochem. biophys. res. commun. 130, 412-417). we report the complete nucleotide sequence of the camc gene along with 155 base pairs of 5' and 175 base pai ...19863003058
[similar organization of beta,beta'-subunit rna-polymerase genes and adjacent ribosomal protein genes in enterobacteriaceae and pseudomonas putida].the genes coding for the rna-polymerase beta,beta'-subunits and adjacent ribosomal protein genes in escherichia coli, salmonella typhimurium, shigella flexneri, serratia marcescens, proteus mirabilis and pseudomonas putida are compared by the southern hybridization procedure. in all the species studied close clustering of the genes rplkajl and rpobc is demonstrated. preliminary physical maps for these genes in s. typhimurium, s. flexneri, s. marcescens and p. mirabilis are proposed. rifampicin i ...19863005845
genetic analysis of a relaxed substrate specificity aromatic ring dioxygenase, toluate 1,2-dioxygenase, encoded by tol plasmid pww0 of pseudomonas putida.toluate 1,2-dioxygenase is the first enzyme of a meta-cleavage pathway for the oxidative catabolism of benzoate and substituted benzoates to krebs cycle intermediates that is specified by tol plasmid pww0 of pseudomonas putida. a collection of derivatives harbouring tn1000 insertions and defective in toluate dioxygenase have been isolated from ppl392, a pbr322-based hybrid plasmid carrying the tol plasmid meta-cleavage pathway operon. in parallel, a series of n-methyl-n'-nitro-n-nitro-soguanidin ...19863010045
analysis of the vegetative replication origin of broad-host-range plasmid rk2 by transposon mutagenesis.a range of tn1723 transposon mutants of the oriv region of broad-host-range plasmid rk2 have been isolated, and the internal ecori fragment of the transposon has been deleted from each to reduce the insertion size from 9.6 kb (tn1723) to 35 bp (delta tn1723). sequencing from the delta tn1723-derived ecori site has allowed the precise mapping of these insertions to various points dispersed through the origin region. using these mutants we have determined which regions of oriv rk2 are of functiona ...19863010353
camr, a negative regulator locus of the cytochrome p-450cam hydroxylase operon.a 4.27-kilobase insert from a hindiii dna library of pseudomonas putida carrying the cam plasmid allowed coordinate expression of genes camd and camc under control of camr, an upstream regulator. the camc gene specifies cytochrome p-450cam, and camd specifies the 5-exo-alcohol dehydrogenase. a 1.38-kilobase deletion from the insert results in the constitutive expression of genes camc and camd; transformation in trans restores the substrate control, indicating that camr is a negative regulator.19863011733
nucleotide sequence of a dna segment promoting transcription in pseudomonas putida.a dna segment that promotes gene expression in pseudomonas putida was identified in ptn8, a mutant plasmid of an rp4-tol recombinant. a promoter on the segment was cloned with a promoter-probe vector containing the xyle gene of the tol plasmid. the xyle gene was expressed under the control of the promoter, and the gene product catechol 2,3-dioxygenase was constitutively synthesized. as analyzed by an s1 nuclease protection assay, the amount of mrna produced in p. putida was more than that in esc ...19863011741
reactions of 3-ethylcatechol and 3-(methylthio)catechol with catechol dioxygenases.the reactions of 3-ethylcatechol and 3-(methylthio)catechol with catechol 1,2-dioxygenase and catechol 2,3-dioxygenase from pseudomonas putida were examined. both 3-substituted catechols are oxidized by catechol 2,3-dioxygenase at approximately 30% of the rate observed for catechol oxidation by this enzyme. analysis of the products of the reactions showed that ring cleavage occurs in a normal fashion between carbons 2 and 3 of the alternate substrates. 3-ethylcatechol is oxidized by catechol 1,2 ...19863015028
characterization of a plasmid-specified pathway for catabolism of isopropylbenzene in pseudomonas putida re204.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 plasmid plg221 yielded mutant derivatives defective in isopropylbenzene metabolism. these were characterized by the identification of the products which they accumulated when grown in the presence of isopropylbenzene and by the assay of enzyme activities in cell extracts. based on the results obtaine ...19863019995
spontaneous deletion of a 20-kilobase dna segment carrying genes specifying isopropylbenzene metabolism in pseudomonas putida re204.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 pre4 of an approximately 20-kilobase segment of dna carrying the catabolic genes. the presence of dna sequences at the ends of the catabolic gene region sharing homology with one another suggests that the deletions result from recombination events between these homologous sequences.19863020004
nucleotide sequence of the regulatory gene xyls on the pseudomonas putida tol plasmid and identification of the protein product.the xyls gene is a regulatory gene which positively controls expression of the genes on the tol plasmid for degradation enzymes of benzoate or m-toluate in pseudomonas putida. cloning of the gene in escherichia coli and determination of the nucleotide sequence revealed an open reading frame of 963 bp which corresponds to a protein with an mr of 36,502. the xyls gene was recloned onto a tac-promoter vector, and the product was identified by the maxicell procedure as a protein with an approximate ...19863023186
[comparative analysis of the organization of the npl-1 plasmid controlling naphthalene oxidation in pseudomonas putida and its derivatives].the paper contains the data on the structure of npl-1 plasmid controlling naphthalene biodegradation. the plasmid which pertains to the p-9 incompatibility group is transferrable conjugatively and is maintained stably within a wide range of gram-negative bacteria. the analysis of mutants and transposon derivatives of the plasmid made it possible to localize nah-genes in a dna fragment, 23 kb in size. an inverted dna segment of 4.2 kb was discovered which may participate in the regulation of nah- ...19863025060
[hybrid plasmid pbs251 containing genes for n-alkane degradation].the strain of pseudomonas aeruginosa bs316 utilizing h-alkanes of the c6-c12 series (alk+) harbours the 96 md plasmid pbs250. the use of plasmid rp4 to mobilize alk+ markers in conjugal transfer to pseudomonas aeruginosa and pseudomonas putida has resulted in isolation of transconjugants resistant to antibiotics (due to genes coded by plasmid rp4) and capable of growth on h-alkanes. a transconjugant from this series harbours plasmid pbs251, a derivative of plasmid rp4 containing the genes for oc ...19853025683
[nah-genes of pseudomonas putida: molecular genetic analysis of the plasmid pbs286].the hybridization and restriction analysis of the plasmid pbs286 (73 kb, the p-9 inc group) as well as parental plasmids npl-1, npl-41 demonstrated that pbs286 plasmid (delta npl-41::tna) with the constitutive synthesis of naphthalene dioxygenase carried genes for naphthalene oxidation to salicylate and those participating in degradation of catechol. restriction map of pbs286 using xhoi restriction endonuclease and that of the nah region using ecori, bamhi, sali and xhoi were established. struct ...19863026897
gene organization of the first catabolic operon of tol plasmid pww53: production of indigo by the xyla gene product.the entire operon coding for the enzymes responsible for conversion of toluenes to benzoates has been cloned from tol plasmid pww53 and the position of the genes accurately located. the coding region was 7.4 kilobase pairs (kbp) long, and the gene order was operator-promoter region (op1)-a small open reading frame-xylc (1.6 kbp)-xyla (2.9 kbp)-xylb (1.8 kbp). within the coding region there was considerable homology with the isofunctional region of the archetypal tol plasmid pww0. a central regio ...19873027047
l-malate transport and proton symport in vesicles prepared from pseudomonas putida.in vesicles from glucose-grown pseudomonas putida, l-malate is transported by nonspecific physical diffusion. l-malate also acts as an electron donor and generates a proton motive force (delta p) of 129 mv which is composed of a membrane potential (delta psi) of 60 mv and a delta ph of 69 mv. in contrast, vesicles from succinate-grown cells transport l-malate by a carrier-mediated system with a km value of 14.3 mm and a vmax of 313 nmol x mg protein-1 x min-1, generate no delta psi, delta ph, or ...19863030368
structure of the pseudomonas putida alkbac operon. identification of transcription and translation products.the structural genes of the pseudomonas oleovorans alk (alkane utilization) system, which are localized on the alkbac operon, were cloned as a 16.9-kilobase pair ecori fragment. we have measured the length and determined the position of the alkbac operon on this fragment by electron microscopy of r-loops. furthermore, the 7.3-kilobase pair long alkbac operon was analyzed for translation products in escherichia coli minicells. using a spectrum of overlapping subclones, six different proteins were ...19873032966
construction of a transposon containing a gene for polygalacturonate trans-eliminase from klebsiella oxytoca.a dna fragment containing a klebsiella oxytoca gene for polygalacturonate trans-eliminase was cloned into the kanamycin resistance transposon tn5. this new transposon, designated tn5-pga+, had a transposition frequency of 1 x 10(-6). the broad host range plasmid pr751::tn5-pga+ was conjugally transferred to a variety of genetic backgrounds. the ability to degrade polygalacturonate was expressed in aeromonas hydrophila, alcaligenes eutrophus, azotomonas insolita, escherichia coli, pseudomonas put ...19873034186
interposon mutagenesis of soil and water bacteria: a family of dna fragments designed for in vitro insertional mutagenesis of gram-negative bacteria.we have constructed a series of derivatives of the omega interposon [prentki and krisch, gene 29 (1984) 303-313] that can be used for in vitro insertional mutagenesis. each of these dna fragments carries a different antibiotic or hg2+ resistance gene (apr, cmr, tcr, kmr or hgr) which is flanked, in inverted orientation, by transcription and translation termination signals and by synthetic polylinkers. the dna of these interposons can be easily purified and then inserted, by in vitro ligation, in ...19873038679
comparison of the amino acid sequences of the transacylase components of branched chain oxoacid dehydrogenase of pseudomonas putida, and the pyruvate and 2-oxoglutarate dehydrogenases of escherichia coli.the nucleotide sequence of bkdb, the structural gene for e2b, the transacylase component of branched-chain-oxoacid dehydrogenase of pseudomonas putida has been determined and translated into its amino acid sequence. the start of bkdb was identified from the n-terminal sequence of e2b isolated from branched-chain-oxoacid dehydrogenase of the closely related species, p. aeruginosa. the reading frame was composed of 65.5% g + c with 82.3% of the codons ending in g or c. there was no intergenic spac ...19883046941
[genes encoding bacterial rna-polymerase. i. cloning of rpobc-operon of pseudomonas putida and its physical map].the p. putida rpobc operon, coding for beta and beta' subunits of rna polymerase, was cloned and its physical map constructed.19883048271
pseudomonas stutzeri ferredoxin: close similarity to azotobacter vinelandii and pseudomonas ovalis ferredoxins.the complete primary structure of pseudomonas stutzeri strain zobell ferredoxin was determined by a combination of protease digestion, edman degradation, and carboxypeptidase digestion and was: tfvvtdncikckytdcvevcpvdcfyegpnflvih pdecidcalcepecpaqaifsedevpedqqefielnadlaevwpnite kkdaladaeewdgvkdklqyler. the calculated molecular weight was 12,110 excluding iron and sulfur atoms. the amino acid sequence was highly homologous to those of azotobacter vinelandii and pseudomonas ovalis ferredoxins. it ...19883053681
purification and properties of carnitine dehydrogenase from pseudomonas putida.carnitine dehydrogenase (carnitine:nad+ oxidoreductase, ec 1.1.1.108) from pseudomonas putida ifp 206 catalyzes the oxidation of l-carnitine to 3-dehydrocarnitine. the enzyme was purified 72-fold to homogeneity as judged by polyacrylamide gel electrophoresis. the molecular mass of this enzyme is 62 kda and consists of two identical subunits. the isoelectric point was found to be 4.7. the carnitine dehydrogenase is specific for l-carnitine and nad+. the optimum ph for enzymatic activity in the ox ...19883058208
[cloning and expression of pseudomonas putida gene controlling the catechol-2,3-oxygenase activity in escherichia coli cells].the genes nahc and nahd from pseudomonas putida naphthalene degradation plasmid pbs286 were cloned on the vector puc19 in escherichia coli cells. the catechol-2,3-oxygenase activity observed in e. coli cells containing recombinant plasmid pbs955 demands the participation of 32 kd polypeptide which is apparently the product of the nahc gene. second polypeptide of molecular weight 34.5 kd is synthesized in pbs955 containing e. coli minicells and perhaps it is a nahd gene product. the data obtained ...19883058550
molecular cloning of 3-phenylcatechol dioxygenase involved in the catabolic pathway of chlorinated biphenyl from pseudomonas putida and its expression in escherichia coli.genes encoding 3-phenylcatechol dioxygenases were cloned from the chlorobiphenyl-degrading pseudomonas putida strain ou83, using broad-host-range cosmid vector pcp13. restriction enzyme analysis of dna from 2,3-dioxygenase-positive chimeric cosmids showed dna inserts ranging in size from 6.0 to 30 kilobases. the origin of the dna insert in hybrid clones was established by using 32p-labeled hybrid clones (poh101 and poh810). a 2.3-kilobase hindiii fragment was common to two clones. the 2,3-dioxyg ...19883063207
[nucleotide sequence of of rpob gene encoding the rna-polymerase beta-subunit in pseudomonas putida]. 19883069413
[nucleotide sequence of the rpoc-gene coding for rna polymerase beta'-subunit of pseudomonas putida]. 19883069416
cloning of genes for branched-chain keto acid dehydrogenase in pseudomonas putida. 19883071713
cytochrome p450: molecular architecture, mechanism, and prospects for rational inhibitor design.cytochromes p450 catalyze the insertion of one o2-derived oxygen atom into an aliphatic or aromatic molecule. p450s are best known for the metabolism of xenobiotic molecules, where hydroxylation renders insoluble hydrocarbons more soluble for easier elimination. in addition to this important catabolic function, p450s catalyze key steps in steroid and plant growth regulator metabolism. a variety of therapeutic, fungicidal, and agochemical agents that perturb these metabolic pathways very likely o ...19883073382
stability fluctuations of plasmid-bearing cells: immobilization effects.the maintenance of the plasmid vectors ptg201 and ptg206 (which both carry the pseudomonas putida xyle gene) and pb lambda h3 in escherichia coli hosts was studied in free and immobilized continuous cultures. ptg201, containing the strong lambda pr promoter, was more quickly lost than plasmid ptg206, containing the tetracycline resistance gene promoter. the instability of ptg201 seems to be related to high expression of the cloned xyle genet. fluctuations in the proportion of ptg201-containing c ...19883075659
cloning and expression of pca genes from pseudomonas putida in escherichia coli.beta-ketoadipate elicits expression of five structural pca genes encoding enzymes that catalyse consecutive reactions in the utilization of protocatechuate by pseudomonas putida. three derivatives of p. putida prs2000 were obtained, each carrying a single copy of tn5 dna inserted into a separate region of the genome and preventing expression of different sets of pca genes. selection of tn5 in or near the pca genes in these derivatives was used to clone four structural pca genes and to enable the ...19883076176
physical and functional mapping of two cointegrate plasmids derived from rp4 and tol plasmid pdk1.cointegrate plasmids were formed in vivo between the broad-host-range r-plasmid rp4 and two catabolic plasmids derived from pseudomonas putida hs1. one of these was the wild-type plasmid pdk1 encoding the complete inducible toluene/xylene (tol) catabolic pathway and one was pdkt1, a deletion derivative of pdk1 selected after growth of hs1 on benzoate and supporting growth on only toluene. the two plasmids formed, pdk2 and pdkt2 respectively, each consisted of a complete rp4 replicon in which was ...19883076182
reported contamination of heparin sodium with pseudomonas putida. 19863080667
amino acid sequence of the pyruvate and the glyoxylate active-site lysine peptide of escherichia coli 2-keto-4-hydroxyglutarate aldolase.pure 2-keto-4-hydroxyglutarate aldolase of escherichia coli, a "lysine-type" trimeric enzyme which has the unique properties of forming an "abortive" schiff-base intermediate with glyoxylate (the aldehydic product/substrate) and of showing strong beta-decarboxylase activity toward oxalacetate, binds any one of its substrates (2-keto-4-hydroxyglutarate, pyruvate, or glyoxylate) in a competitive manner. to determine whether the substrates bind at the same or different (juxta-positioned) sites and ...19863090043
utilization of pyrimidines and pyrimidine analogues by fluorescent pseudomonads.the fluorescent pseudomonads pseudomonas aeruginosa, pseudomonas aureofaciens and pseudomonas putida were examined for their ability to utilize pyrimidines and pyrimidine analogues. both p. aeruginosa and p. aureofaciens grew upon dihydrouracil, dihydrothymine, uridine and cytidine as either a sole nitrogen or carbon source while uracil, thymine and cytosine served only as sole nitrogen sources for both pseudomonads. the only difference between the observed growth of p. aeruginosa and p. aureofa ...19863097460
a comparative study of acquired amidase activity in pseudomonas species.pseudomonas putida pp3 carrying dehalogenases i and ii and pseudomonas aeruginosa pau3 carrying dehalogenase i coded for by plasmid puu2 were able to grow on 2-monochloropropionic acid (2mcpa). neither strain utilized 2-chloropropionamide (2cpa) as a carbon or nitrogen source for growth. mutations in both strains to 2cpa+ phenotypes (designated p. putida ppw3 and p. aeruginosa pau5, respectively) involved the expression of an acquired 2cpa-amidase activity. the amidase followed by dehalogenase r ...19863098906
purification and characterization of glyoxalase i from pseudomonas putida.glyoxalase i was purified to apparent homogeneity from pseudomonas putida. the enzyme was a monomer with a molecular weight of 20,000. the enzyme was most active at ph 8.0. the km values for methylglyoxal and 4,5-dioxovale-rate are 3.5 mm and 1.2 mm, respectively. contrary to the case of eukaryotic enzymes, chelating agents showed little inhibitory effects on the enzyme activity. among the metal ions tested, zn++ specifically and completely inhibited the activity of the enzyme at a millimolar le ...19863101683
the amino acid sequence of the aliphatic amidase from pseudomonas aeruginosa.amino acid sequence studies show that the aliphatic amidase (ec 3.5.1.4) from pseudomonas aeruginosa pac142 consists of a single polypeptide chain of 346 residues, giving an mr of 38,400. the evidence from the amino acid studies is in complete agreement with that deduced from the dna sequence of the amie gene. studies of the protein from pseudomonas putida a87 show that it differs from the ps. aeruginosa protein by about 30 amino acid substitutions. it now becomes possible to relate changes in t ...19873108029
genetic analysis of mannityl opine catabolism in octopine-type agrobacterium tumefaciens strain 15955.the genetic organization of functions responsible for mannityl opine catabolism of the ti plasmid of agrobacterium tumefaciens strain 1,5955 was investigated. a partial hindiii digest of pti1,5955 was cloned into a broad host range cosmid and the clones obtained were tested for ability to confer mannityl opine degradation upon agrobacterium. inserts containing genes for catabolism of mannopinic acid, mannopine, agropine, and agropinic acid were obtained, spanning a segment of 43 kb on the ti pla ...19873112522
[automated micromethod for the determination of the utilization of carbon sources by clinically significant pseudomonas species].the assimilation of 43 different carbon substrates by 93 clinical strains of pseudomonas aeruginosa was studied by a new miniaturized rapid method. reading of assimilation results was done photometrically after 18-20 h incubation and the resulting data were captured and stored by a microcomputer. the differentiating capacity of the assimilation tests were verified by comparing the results of 41 strains of pseudomonas fluorescens, 48 strains of pseudomonas putida, 52 strains of pseudomonas maltop ...19873118596
[expression of the human interferon alpha f gene in the obligate methylotroph methylobacillus flagellatum kt and pseudomonas putida].the expression of human leucocyte interferon alpha f gene in plasmid plm-ifn alpha f-273 is controlled by a hybrid tac (trp-lac) promoter. a structural gene for interferon alpha f is a component of the hybrid operon lacz'-ifn alpha f-tcr, that contains an e. coli trp-operon intercystronic region. plasmid plm ifn alpha f-273--directed interferon synthesis allows to obtain about 10(7) iu/l. this plasmid was cloned in broad-host-range vector plasmid payc31. the hybrid bi-repliconed plasmid containi ...19873119998
a set of cassettes and improved vectors for genetic and biochemical characterization of pseudomonas genes.a set of broad-host-range vectors allowing direct selection of recombinant dna molecules to facilitate subcloning and expression analyses of pseudomonas genes was constructed using bg/ii lacz alpha cassette. controlled expression vectors pvdtac39 and pvdtac24 were shown to be useful for determination of enzymatic activities encoded by the cloned dna fragments and mr determination of the corresponding polypeptides. a set of pseudomonas putida xyle gene cassettes truncated at the 5' end was constr ...19873123327
[isolation and comparative characteristics of 2 unrelated temperate phages of pseudomonas putida ppg1].two temperate bacteriophages, pp56 and pp71, specific for bacteria of pseudomonas putida strain ppg1 have been isolated for the first time. characterization of the phages was performed. both of them accomplish stable lysogenization of p. putida ppg1 cells. the phages are inducible. several groups of clear plaque (c) mutants of pp56 and pp71 with altered processes of establishment and maintenance of lysogenic state have been isolated, according to complementation test. the phages differ in follow ...19883129337
purification and characterization of cutinase from a fluorescent pseudomonas putida bacterial strain isolated from phyllosphere.cutinase, an extracellular enzyme, was induced by cutin in a fluorescent pseudomonas putida strain that was found to be cohabiting with an apparently nitrogen-fixing corynebacterium. this enzyme was purified from the culture fluid by acetone precipitation followed by chromatography on deae-cellulose, qae-sephadex, sepharose 6b, and sephadex g-100. the purified enzyme showed a single band when subjected to polyacrylamide electrophoresis and the enzymatic activity coincided with the protein band. ...19883130804
physiological comparison of d-cysteine desulfhydrase of escherichia coli with 3-chloro-d-alanine dehydrochlorinase of pseudomonas putida cr 1-1.d-cysteine desulfhydrase of escherichia coli w3110 delta trped102/f' delta trped102 was physiologically characterized. it was found to be located in the cytosolic fraction, as 3-chloro-d-alanine dehydrochlorinase is. d-cysteine desulfhydrase catalyzed not only the alpha, beta-elimination reaction of o-acetyl-d-serine to form pyruvate, acetic acid and ammonia, but also the beta-replacement reaction of o-acetyl-d-serine with sulfide to form d-cysteine. however, these reactions appeared not to proc ...19883132906
transformation of bacteria with plasmid dna by electroporation.the possibility of electric field-mediated transformation ("electroporation") of a gram-positive bacterium (enterococcus faecalis) and two gram-negative bacteria (escherichia coli and pseudomonas putida) with plasmid dna was investigated. e. faecalis protoplasts could be transformed by electroporation with a transformation frequency of 10(4) to 10(5) transformants/micrograms plasmid. untreated--i.e., washed--cells of e. coli could be transformed with rates of 1 x 10(5) transformants/micrograms p ...19883133958
the complete amino acid sequence and identification of the active-site arginine peptide of escherichia coli 2-keto-4-hydroxyglutarate aldolase.the complete amino acid sequence of 2-keto-4-hydroxyglutarate aldolase from escherichia coli has been established in the following manner. after being reduced with dithiothreitol, the purified aldolase was alkylated with iodoacetamide and subsequently digested with trypsin. the resulting 19 peptide peaks observed by high performance liquid chromatography, which compared with 21 expected tryptic cleavage products, were all isolated, purified, and individually sequenced. overlap peptides were obta ...19883136164
[bacteriophages of pseudomonas putida containing single-stranded canonical dna breaks].it was shown that bacteriophage tf as well as bacteriophages phi p4/40, phi p25/42, phi p23/40 and phi p6/40, which are specific to different p. putida strains, contain the single strand breaks in their dna. the breaks are localized in one strand of dna molecules and are repairable with t4 dna ligase. bacteriophage tf has no detectable dna homology with phi p4/40, phi p25/42, phi p23/40 and phi p6/40 bacteriophages. all the phages studied have no relation with other known pseudomonas phages. bac ...19883137462
cloning and expression of the cata and catbc gene clusters from pseudomonas aeruginosa pao.a 9.9-kilobase (kb) bamhi restriction endonuclease fragment encoding the cata and catbc gene clusters was selected from a gene bank of the pseudomonas aeruginosa pao1c chromosome. the cata, catb, and catc genes encode enzymes that catalyze consecutive reactions in the catechol branch of the beta-ketoadipate pathway: cata, catechol-1,2-dioxygenase (ec 1.13.11.1); catb, muconate lactonizing enzyme (ec 5.5.1.1); and catc, muconolactone isomerase (ec 5.3.3.4). a recombinant plasmid, pro1783, which c ...19883139626
construction of a shuttle vector for inducible gene expression in escherichia coli and bacillus subtilis.the construction of a shuttle vector for inducible gene expression allowing fast and easy cloning in escherichia coli and subsequent transformation of bacillus subtilis is presented. the expression is based on the regulation of the tac promoter by the lac repressor which was assayed with the xyle gene from pseudomonas putida as a marker gene. the laciq gene, transcribed by the strong spo promoter, allowed full repression of the weak tac promoter.19883141570
survival of rifampin-resistant mutants of pseudomonas fluorescens and pseudomonas putida in soil systems.the fate of spontaneous chromosomal rifampin-resistant (rifr) mutants of pseudomonas putida and pseudomonas fluorescens in sterile and live organic soil from which they were isolated was studied. in sterile native-soil assays, a rifr mutant of p. putida showed no decrease in competitive fitness when compared with the wild-type parent. however, mutants of p. fluorescens were of two general categories. group 1 showed no difference from the wild type in terms of growth rate, competitive fitness, an ...19883144244
mineralization of the dibenzothiophene biodegradation products 3-hydroxy-2-formyl benzothiophene and dibenzothiophene sulfone.dibenzothiophene is degraded to 3-hydroxy-2-formyl benzothiophene by various bacteria, including a strain of pseudomonas putida that also forms dibenzothiophene sulfone via an alternate pathway. by using these end products as substrates, mixed enrichment cultures that could degrade 3-hydroxy-2-formyl benzothiophene and dibenzothiophene sulfone with the formation of co2 were established.19883146950
minimum bacterial density for bacteriophage replication: implications for significance of bacteriophages in natural ecosystems.bacteriophage 80 alpha did not increase in number in cultures containing less than about 1.0 x 10(4) to 1.5 x 10(4) cfu of staphylococcus aureus per ml, but bacteriophage replication did occur when the number of bacteria exceeded this density, either initially or as a result of host cell multiplication. the minimum density of an asporogenous strain of bacillus subtilis required for an increase in the number of bacteriophage sp beta ci was about 3 x 10(4) cfu/ml. the threshold density of escheric ...19853156556
nucleotide sequence of the regulatory gene xylr of the tol plasmid from pseudomonas putida.we have determined the nucleotide sequence of the xylr gene for a transcriptional activator for the degradative pathway of aromatic hydrocarbons on the tol plasmid from pseudomonas putida. the 1698-bp sequence for a 566-amino acid (aa) protein (mr 63741) was identified as the xylr-encoding sequence. three regions in xylr show homology to klebsiella pneumoniae ntrc and nifa, both of which are transcriptional activators for the ntr and nif genes involved in the nitrogen metabolism. the central reg ...19883169574
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