Publications

transposable elements for efficient manipulation of a wide range of gram-negative bacteria: promoter probes and vectors for foreign genes.	we describe here the construction and use of a series of modified transposons based on the insertion sequence is1. like their parent, omegon-km [fellay et al., gene 76 (1989) 215-226], these elements permit efficient insertional mutagenesis of a variety of gram-negative bacteria. the presence of a functional pbr322 origin of replication within the transposable element facilitates subsequent cloning of the mutated gene. the omegon-km system was previously shown to function in pseudomonas putida, ...	1989	2559879
[cloning and gene expression determining phenol breakdown in pseudomonas putida strains].		1989	2561419
study of the 5-oxoprolinase reaction by 13c nmr.	5-oxoprolinase catalyzes the atp-dependent decyclization of 5-oxo-l-proline to l-glutamate. previous studies provided evidence for the intermediate formation of a phosphorylated form of 5-oxoproline (seddon, a. p., and meister, a. (1986) j. biol. chem. 261, 11538-11541) and of a tetrahedral intermediate (li, l., seddon, a. p., and meister, a. (1987) j. biol. chem. 262, 11020-11025). a new approach to the study of the reaction mechanism using the 18o isotope effect on the 13c nmr signals for 5-ox ...	1989	2563377
thermal activation of photoactivatable urocanase from pseudomonas putida.	the dark inactivation of urocanase from pseudomonas putida is caused by the formation of a sulfite adduct of the tightly bound coenzyme, nicotinamide adenine dinucleotide. photodissociation of this adduct by uv radiation restores the enzyme activity. based on cold exhaustive dialysis the modification reaction appeared to be irreversible. however, we now report that sulfite modification of urocanase is reversible at higher temperatures. an arrhenius plot of the thermal activation is linear (20-38 ...	1989	2570140
the stoichiometry of the tightly bound nad+ in urocanase. separation and characterization of fully active and inhibited forms of the enzyme.	1. urocanase, purified by classical methods [keul, v., kaeppeli, f., ghosh, c., krebs, t., robinson, j. a. and rétey, j. (1979) j. biol. chem. 254, 843-851] from pseudomonas putida was submitted to high-performance liquid chromatography on a tsk-deae column. the enzyme was eluted in three resolved peaks (a, b and c) exhibiting specific activities of 3.4 u/mg, 1.85 u/mg and 0.4 u/mg, respectively. 2. the difference spectra of peaks b and a as well as of c and a showed maxima at 330 nm. 3. irradia ...	1989	2574107
specific inhibition of bacterial and bovine urocanases by glycylglycine.	urocanase (ec 4.2.1.49) purified from pseudomonas putida was unexpectedly inhibited by the dipeptide glycylglycine. using a spectrophotometric assay for urocanase activity, we characterized the inhibition. the inhibition was temperature-, concentration-, and time-dependent; 0.1, 0.5 and 1.0 mm glycylglycine inhibited the enzyme by 20%, 50% and 78%, respectively, in 60 min at 30 degrees c. dithiothreitol and reduced glutathione did not prevent the process. the inhibition was a pseudo first-order ...	1989	2577699
operon structure and nucleotide homology of the chlorocatechol oxidation genes of plasmids pjp4 and pac27.	alcaligenes eutrophus harboring plasmid pjp4 (strain jmp134) is capable of growing on both 2,4-dichlorophenoxyacetate (2,4-d) and 3-chlorobenzoate (3-cba), while pseudomonas putida carrying plasmid pac27 (strain ac867) can utilize only 3-cba as the sole carbon source. the tfdcdef operon of the pjp4 plasmid and the clcabd operon of plasmid pac27 each encode enzymes for the degradation of chlorocatechols (clc), key intermediates in the catabolism of 2,4-d and 3-cba. similarities in the nucleotide ...	1989	2583528
survival of and plasmid stability in pseudomonas and klebsiella spp. introduced into agricultural drainage water.	cell survival and plasmid stability in pseudomonas fluorescens r2f and pseudomonas putida cym 318 containing respectively, plasmid rp4 and prk2501, and klebsiella aerogenes nctc 418 harboring plasmid pbr322 were studied in sterile and nonsterile agricultural drainage water under both aerobic and anaerobic conditions and in the absence and presence of added nutrients. both pseudomonas strains survived well in sterile drainage water incubated aerobically, with or without added nutrients. however, ...	1989	2590305
xyle functions as an efficient reporter gene in streptomyces spp.: use for the study of galp1, a catabolite-controlled promoter.	we describe the development of a convenient and sensitive reporter gene system for streptomyces spp. based on the use of a promoterless copy of the xyle gene of pseudomonas putida. the xyle gene product is a catechol dioxygenase, which converts the colorless substrate catechol to an intensely yellow hydroxymuconic semialdehyde. a promoterless copy of xyle was placed under the transcriptional control of galp1, a glucose-repressed and galactose-induced promoter from streptomyces lividans, and its ...	1989	2592344
molecular cloning, coding nucleotides and the deduced amino acid sequence of p-450bm-1 from bacillus megaterium.	the gene encoding barbiturate-inducible cytochrome p-450bm-1 from bacillus megaterium atcc 14581 has been cloned and sequenced. an open reading frame in the 1.9 kb of cloned dna correctly predicted the nh2-terminal sequence of p-450bm-1 previously determined by protein sequencing, and, in toto, predicted a polypeptide of 410 amino acid residues with an mr of 47,439. the sequence is most, but less than 27%, similar to that of p-450cam from pseudomonas putida, so that p-450bm-1 clearly belongs to ...	1989	2597681
[study of morphology and genome structure of pseudomonas putida bacteriophages for their classification].	a group of 27 bacteriophages specific for pseudomonas putida strains ppg1 and ppn has been isolated. the phages were characterized and compared with the previously described virulent (pf 16, af, tf and pmw) and temperate (pp56 and pp71) phages. the new phages belong to b1 and c1 morphotypes, according to ackerman's classification. phage dnas were digested with several endonucleases; the molecular weights and homology of the dnas were determined. all phages of p. putida isolated up to now were di ...	1989	2599372
nucleotide and deduced amino acid sequence of the rpon sigma-factor of pseudomonas putida.		1989	2602128
survival of pseudomonas putida uwc1 containing cloned catabolic genes in a model activated-sludge unit.	the possibility of the accidental or deliberate release of genetically engineered microorganisms into the environment has accentuated the need to study their survival in, and effect on, natural habitats. in this study, pseudomonas putida uwc1 harboring a non-self-transmissible plasmid, pd10, encoding the breakdown of 3-chlorobenzoate was shown to survive in a fully functioning laboratory-scale activated-sludge unit (asu) for more than 8 weeks. the asu maintained a healthy, diverse protozoal popu ...	1989	2604401
monohydroxylation of phenol and 2,5-dichlorophenol by toluene dioxygenase in pseudomonas putida f1.	pseudomonas putida f1 contains a multicomponent enzyme system, toluene dioxygenase, that converts toluene and a variety of substituted benzenes to cis-dihydrodiols by the addition of one molecule of molecular oxygen. toluene-grown cells of p. putida f1 also catalyze the monohydroxylation of phenols to the corresponding catechols by an unknown mechanism. respirometric studies with washed cells revealed similar enzyme induction patterns in cells grown on toluene or phenol. induction of toluene dio ...	1989	2604403
cloning and nucleotide sequences of nadh-putidaredoxin reductase gene (cama) and putidaredoxin gene (camb) involved in cytochrome p-450cam hydroxylase of pseudomonas putida.	pseudomonas putida ppgl, which carries the cam plasmid encoding enzymes involved in the degradation pathway of d-camphor, can utilize d-camphor as a sole carbon source. cytochrome p-450cam and related enzymes participate in the early oxidation steps of d-camphor degradation metabolism. we cloned from a hindiii dna library of ppgl a 2.9 kbp cam segment which carries the major part of cama gene encoding nadh-putidaredoxin reductase and the entire camb gene encoding putidaredoxin. the 2.9 kbp cam s ...	1989	2613690
[plasmids for biphenyl, chlorobiphenyl and metatoluylate degradation from pseudomonas putida].	pseudomonas putida strain su83, harbors the pbs311 plasmid coding for the degradation of biphenyl, 2- and 4-chlorbiphenyl, meta- and paratoluylate. the insertional mutants of the plasmid obtained by the transposon tn5 insertion were isolated. one of the mutants was used for cloning of the biphenyl degradation genes. the plasmid pbs311:: tn5 dna was inserted into the bamhi site of the plasmid pbr322 and cloned. 11 recombinants of 354 tested were treated with 0.1% solution of 2,3-dioxybiphenyl. on ...	1989	2628753
[genetic determination of degradation of ampholytic surfactants].	plasmid dna was detected in pseudomonas putida 141 and p. stutzeri at strains which caused destruction of the ampholytic surfactants alkylamino-bis-propionate (aabp) and amidobetaine, respectively. as was demonstrated using genetic analytic procedures, the plasmids controlled aabp and amidobetaine destruction. no plasmid dna was found in p. desmolytica c37 which caused cyclimide destruction or in pseudomonas sp. 1 and citrobacter freundii to strains responsible for aabp destruction. apparently, ...	1989	2636974
extracellular product of nocardia amarae induces bacterial cell flocculation.	the fact that nocardia amarae yk1 produced a bacterial flocculation-inducing substance (designated as fix) was discovered. fix had a function of flocculating proliferous cells. fix-induced flocculation was inhibited by making cells resting, but not completely by adding chloramphenicol. fix worked widely on gram-positive to -negative bacteria. in the presence of fix, achromobacter cycloclastus iam1013, acinetobacter calcoaceticus iam1517, bacillus subtilis iam1069, escherichia coli c600-1, e. col ...	1989	2653957
in vivo enzymology: a deuterium nmr study of formaldehyde dismutase in pseudomonas putida f61a and staphylococcus aureus.	high-resolution deuterium nmr spectroscopy has been used to follow the detoxifying metabolism of [d2]formaldehyde in vivo in several bacterial species. production of [d2]methanol in escherichia coli confirms that the oxidation and reduction pathways of metabolism are independent in this organism. efficient production of equimolar quantities of [d]formate and [d3]methanol in pseudomonas putida f61a and staphylococcus aureus implicates a formaldehyde dismutase, or "cannizzarase", activity. these o ...	1989	2655705
cloning and expression in escherichia coli of the toluene dioxygenase gene from pseudomonas putida ncib11767.	the genes encoding toluene dioxygenase, toluene cis-glycol dehydrogenase and catechol 2.3-oxygenase from pseudomonas putida ncib 11767 were cloned and expressed in escherichia coli hb101 on a 20 kb fragment. the recombinant strain produced indigo and a variety of other coloured products. although the enzymes were expressed in the absence of inducers, further induction was observed in the presence of toluene or benzene, implying the presence of regulatory elements on the 20 kb insert.	1989	2656389
5-carboxymethyl-2-hydroxymuconic semialdehyde dehydrogenases of escherichia coli c and klebsiella pneumoniae m5a1 show very high n-terminal sequence homology.	5-carboxymethyl-2-hydroxymuconic semialdehyde (chms) dehydrogenase from escherichia coli c and klebsiella pneumoniae m5a1 have been purified and some of their properties studied. the apparent km values for nad and chms were 11.7 +/- 1.5 microm and 5.2 +/- 1.9 microm, respectively, for the k. pneumoniae enzyme, and 19.5 +/- 2.7 microm and 9.2 +/- 1.4 microm, respectively, for the e. coli enzyme. both enzymes were optimally active at ph 7.5 in sodium phosphate buffer. they had subunit molecular we ...	1989	2656390
[cloning of pseudomonas putida genes responsible for the primary stages of oxidation of naphthalene in escherichia coli cells].	data on cloning pseudomonas putida d-plasmid pbs286 (incp-9) genes which are responsible for primary stages of naphthalene oxidation as well as data on the expression of these genes in escherichia coli cells are presented. recombinant plasmid pbs959 containing the whole constitutive naha locus encoding naphthalene dioxygenase, a key enzyme of the naphthalene oxidation pathway, has been constructed on the basis of the puc19 vector. an evidence has been obtained that at least a portion of the sequ ...	1989	2661326
cloning of a carbofuran hydrolase gene from achromobacter sp. strain wm111 and its expression in gram-negative bacteria.	a 14-kilobase-pair (kbp) ecori dna fragment that encodes an enzyme capable of rapid hydrolysis of n-methylcarbamate insecticides (carbofuran hydrolase) was cloned from carbofuran-degrading achromobacter sp. strain wm111. when used to probe southern blots containing plasmid and total dnas from wm111, this 14-kbp fragment hybridized strongly to a 14-kbp ecori fragment from the greater than 100-kbp plasmid harbored by this strain but weakly to ecori-digested total dna from achromobacter sp. strain ...	1989	2661544
2-oxoaldehyde metabolism in microorganisms.	the properties of methylglyoxal-metabolizing enzymes in prokaryotic and eukaryotic microorganisms were studied systematically and compared with those of mammalian enzymes. the enzymes constitute a glycolytic bypass and convert methylglyoxal into pyruvate via lactate. the first step in this conversion is catalyzed by glyoxalase i, methylglyoxal reductase, or methylglyoxal dehydrogenase. the regulation of the yeast glyoxalase system was analyzed. the system was closely related to the proliferative ...	1989	2663129
[susceptibilities of clinical isolates to antibacterial agents. a study mainly focused on ofloxacin (the second report). reported by the research group for testing ofloxacin susceptibility on clinical isolates].	susceptibilities of various clinical isolates to ofloxacin (oflx) and other antibacterial drugs were examined at 128 hospital laboratories in 36 prefectures throughout japan between april, 1986 and march, 1987. the results were totalized with an emphasis mainly on oflx and were compared with data obtained in the previous year. in this study, identification and susceptibility tests of the isolates were carried out at each hospital laboratory and the tests were performed according to the 1-dilutio ...	1989	2664255
identification of multiple repressor recognition sites in the hut system of pseudomonas putida.	the hutc gene in pseudomonas putida encodes a repressor protein that negatively regulates the expression of all hut genes. we have overexpressed this cloned hutc gene in escherichia coli to identify p. putida hut regions that could specifically bind the repressor. ten restriction fragments, some of which were partially overlapping and spanned the coding portions of the p. putida hut region, were labeled and tested for their ability to recognize repressor in a filter binding assay. this procedure ...	1989	2666390
involvement of pseudomonas putida rpon sigma factor in regulation of various metabolic functions.	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 gene and identified its gene product as a protein with an apparent molecular weight of 78,000. a mutant rpon gene was constructed by in vitro insertion mutagenesis with a kanamycin cassette. a p. putida rpon mutant was then isolated by replacement of the intact chromosomal rpon gene by the mutant rpo ...	1989	2666396
differences between the manganese- and the iron-containing superoxide dismutases of escherichia coli detected through sedimentation equilibrium, hydrodynamic, and spectroscopic studies.	the genome of escherichia coli codes for two superoxide dismutases that may contain either iron (fesod) or manganese (mnsod) at the active site. the crystal structures of mnsods from two bacterial sources (but not e. coli) have been completed, and structural comparisons with the crystal structure of the fesod from either e. coli or pseudomonas ovalis have been made. despite the low degree (less than 50%) of sequence homology between the e. coli enzymes, the two proteins are suggested to be struc ...	1989	2669953
transcription initiation at multiple promoters of the pfl gene by e sigma 70-dependent transcription in vitro and heterologous expression in pseudomonas putida in vivo.	in vitro transcription experiments were used to provide further evidence that the gene encoding pyruvate formate-lyase (ec 2.3.1.54) from escherichia coli is transcribed from seven promoters which cover a region of 1.2 kilobase pairs of dna (g. sawers and a. böck, j. bacteriol., 171:2485-2498, 1989). the results demonstrated that all promoters were recognized by the major rna polymerase holoenzyme species e sigma 70 in vitro. further corroboration for multiple functional promoters came from hete ...	1989	2670899
toluene degradation by pseudomonas putida f1. nucleotide sequence of the todc1c2bade genes and their expression in escherichia coli.	the nucleotide sequence of the todc1c2bade genes which encode the first three enzymes in the catabolism of toluene by pseudomonas putida f1 was determined. the genes encode the three components of the toluene dioxygenase enzyme system: reductasetol (toda), ferredoxintol (todb), and the two subunits of the terminal dioxygenase (todc1c2); (+)-cis-(1s, 2r)-dihydroxy-3-methylcyclohexa-3,5-diene dehydrogenase (todd); and 3-methylcatechol 2,3-dioxygenase (tode). knowledge of the nucleotide sequence of ...	1989	2670929
bacterial aromatic ring-cleavage enzymes are classified into two different gene families.	dioxygenases that catalyze the cleavage of the aromatic ring are classified into two groups according to their mode of ring fission. substrates of ring-cleavage dioxygenases usually contain hydroxyl groups on adjacent aromatic carbons, and intradiol enzymes cleave the ring between these two hydroxyl groups. extradiol enzymes in contrast cleave the ring between one hydroxylated carbon and its adjacent nonhydroxylated carbon. in this study, we determined the complete nucleotide sequence of nahc, t ...	1989	2670937
physically associated enzymes produce and metabolize 2-hydroxy-2,4-dienoate, a chemically unstable intermediate formed in catechol metabolism via meta cleavage in pseudomonas putida.	the meta-cleavage pathway of catechol is a major mechanism for degradation of aromatic compounds. in this pathway, the aromatic ring of catechol is cleaved by catechol 2,3-dioxygenase and its product, 2-hydroxymuconic semialdehyde, is further metabolized by either a hydrolytic or dehydrogenative route. in the dehydrogenative route, 2-hydroxymuconic semialdehyde is oxidized to the enol form of 4-oxalocrotonate by a dehydrogenase and then further metabolized to acetaldehyde and pyruvate by the act ...	1989	2681159
isolation and characterization of altered plasmids in mutant strains of pseudomonas putida ncib 9816.	the ability of p. putida ncib 9816 to grow with naphthalene (nah+) and salicylate (sal+) is correlated with the presence of an 83 kilobase (kb) conjugative plasmid, pdtg1. derivatives of pdtg1 were obtained from cells after exposure to halogenated analogs of naphthalene or salicylate. the selection of mutants having a nah-sal- or a nah-sal+ phenotype could be enhanced by the addition of triphenyltetrazolium chloride to the indicator medium. structurally modified plasmids were characterized by re ...	1989	2684156
putidaredoxin competitively inhibits cytochrome b5-cytochrome p-450cam association: a proposed molecular model for a cytochrome p-450cam electron-transfer complex.	cytochrome b5 has been genetically engineered to afford a fluorescent derivative capable of monitoring its association with cytochrome p-450cam from pseudomonas putida [stayton, p. s., fisher, m. t., & sligar, s. g. (1988) j. biol. chem. 263, 13544-13548]. in the mutant cytochrome b5, threonine is replaced by a cysteine at position 65 (t65c) and has been labeled with the environmentally sensitive fluorophore acrylodan. in this paper, the physiological p-450cam reductant putidaredoxin, an fe2s2.c ...	1989	2690937
trichloroethylene degradation by escherichia coli containing the cloned pseudomonas putida f1 toluene dioxygenase genes.	toluene dioxygenase from pseudomonas putida f1 has been implicated as an enzyme capable of degrading trichloroethylene. this has now been confirmed with escherichia coli jm109(pdtg601) that contains the structural genes (todc1c2ba) of toluene dioxygenase under the control of the tac promoter. the extent of trichloroethylene degradation by the recombinant organism depended on the cell concentration and the concentration of trichloroethylene. a linear rate of trichloroethylene degradation was obse ...	1989	2694960
microbial enzymes for creatinine assay: a review.	a novel metabolic pathway for the degradation of creatinine with n-methylhydantoin, n-carbamoylsarcosine and sarcosine as successive intermediates was found to operate in pseudomonas putida 77 and many other microorganisms. enzymes involved in this pathway were purified from cells of p. putida 77 and characterized. the first step, deimination of creatinine, is catalyzed by cytosine deaminase/creatinine deiminase. the following two steps, ring-opening of n-methylhydantoin and decarbamoylation of ...	1989	2695273
cloning and sequence analysis of the ntra (rpon) gene of pseudomonas putida.	the gene encoding a sigma factor ntra (rpon) was cloned from pseudomonas putida by cross-hybridization with a probe containing a part of the corresponding escherichia coli gene. the cloned gene complemented an ntra mutation of e. coli in activation of xyl genes on the tol plasmid. the predicted amino acid (aa) sequence of p. putida ntra (497 aa; mr 56,215) is highly homologous to ntra proteins from azotobacter vinelandii (81.7%), klebsiella pneumoniae (52.6%), and rhizobium meliloti (36.1%). the ...	1989	2695395
mutations in genes downstream of the rpon gene (encoding sigma 54) of klebsiella pneumoniae affect expression from sigma 54-dependent promoters.	two open reading frames (orfs), designated orf95 and orf162, downstream of the klebsiella pneumoniae sigma 54 structural gene (rpon) have been sequenced and shown to encode polypeptides of 12 kd and 16 kd, respectively. orfs homologous to orf95 are present downstream of four out of five rpon genes sequenced to date from a range of gram-negative bacteria, and orf162 is also conserved, at least in pseudomonas putida. chromosomal mutations have been created in each gene using a kan cassette and bot ...	1989	2695747
evidence that the transcription activator encoded by the pseudomonas putida nahr gene is evolutionarily related to the transcription activators encoded by the rhizobium nodd genes.	the nahr gene of the 83-kilobase naphthalene degradation plasmid nah7 of pseudomonas putida encodes a 34-kilodalton polypeptide which binds to the nah and sal promoters to activate transcription of the degradation genes in response to the inducer salicylate. the dna sequence of the nahr gene was determined, and a derived amino acid sequence of the nahr protein was obtained. a computer search for homologous proteins showed that within the first 124 amino-terminal residues, nahr has approximately ...	1989	2703465
construction and nucleotide sequence of a cdna encoding the full-length preprotein for human branched chain acyltransferase.	a cdna (1.6 kilobases) for branched chain acyltransferase (e2b) isolated from a human liver library encoded only the amino-terminal half of the protein (hummel, k. b., litwer, s., bradford, a. p., aitken, a., danner, d. j., and yeaman, s. j. (1988) j. biol. chem. 263, 6165-6168). here we report the isolation of other cdnas which encode the carboxyl-terminal half of e2b and the construction of a cdna which encodes the entire pre-e2b. cdna from the original clone encoding the leader sequence, lipo ...	1989	2708389
nad-linked, gsh- and factor-independent aldehyde dehydrogenase of the methylotrophic bacterium, hyphomicrobium x.	cell-free extracts of hyphomicrobium x showed nad-dependent aldehyde dehydrogenase activity, provided that nad addition preceded that of aldehyde. activity was lost rather rapidly, especially during purification attempts, but this could be partially masked by including a time-dependent restoration step with thiol compounds in the protocol. the nature of the assay buffer appeared to be critical and stimulation occurred on incorporation of k+ ions in the mixture. an even higher specific activity c ...	1989	2712573
identification of pseudomonas alcaligenes chromosomal dna in the plasmid dna of the chlorobenzene-degrading recombinant pseudomonas putida strain cb1-9.	the recombinant pseudomonas putida strain cb1-9, which acquired the ability to grow on chlorobenzenes, contains a 33-kilobase (kb) plasmid (pkfl3) which lacked homology to an indigenous 15-kb plasmid (pkfl1) in pseudomonas alcaligenes c-0 parent but was homologous to a 55-kb plasmid (pkfl2) from the p. putida r5-3 parent. chromosomal dna of p. alcaligenes c-0 hybridized to probes prepared from pkfl3 but not to probes prepared from pkfl2. a single clone from a genomic library of p. alcaligenes c- ...	1989	2729978
cloning of bacterial genes specifying degradation of 4-chlorobiphenyl from pseudomonas putida ou83.	genes capable of 4-chlorobiphenyl (4-cbp) degradation were cloned from 4-cbp-degrading pseudomonas putida ou83 by using a genomic library which was constructed in the broad-host-range cosmid vector pcp13. p. putida ac812 containing chimeric cosmid-expressing enzymes involved in the 4-cbp degradation pathway were identified by detecting 3-phenylcatechol dioxygenase activity (3-pda). chimeric cosmid clones poh83, poh84, poh85, poh87, and poh88 positive for 3-pda grew in synthetic basal medium cont ...	1989	2729981
[production of antibiotic substances by natural variants of the marine bacterium vibrio fischeri].	it was shown that under definite conditions there was competition between natural variants of sea bacteria belonging to v. fischeri. natural variants of v. fischeri, strain 6 differed in their resistance to streptomycin and had different growth rates under conditions of limited aeration. morphologically all the variants were identical. v. fischeri p-0, v. fischeri p-1 and v. fischeri p-2 were studied. the study revealed that v. fischeri p-0 produced a non-dialysing thermostable trypsin-sensitive ...	1989	2730220
flagellation of pseudomonas putida and analysis of its motile behavior.	pseudomonas putida flagella were examined. also, changes in motile behavior in response to chemoattractants were analyzed quantitatively by computer. reversals in the rotation direction of bundles of polar flagella resulted in changes in swimming direction. cells swimming in buffer changed direction once every 2 s on average, whereas cells exposed to the attractant benzoate changed direction an average of once every 10 s. the findings show that p. putida responds to temporal gradients of chemoat ...	1989	2738028
[nucleotide sequence of the rpll gene coding for ribosomal protein l7/l12 of pseudomonas putida].	the pseudomonas putida rpl l gene coding for ribosomal protein l7/l12 was cloned and sequenced. although asp55 residue in l7/l12 was previously shown to be conservative in ten different organisms, the pseudomonas putida l7/l12 proved to contain asn55, thus showing that asp55 is not invariant.	1989	2751714
a methyl-accepting protein is involved in benzoate taxis in pseudomonas putida.	pseudomonas putida is attracted to at least two groups of aromatic acids: a benzoate group and a benzoylformate group. members of the benzoate group of chemoattractants stimulated the methylation of a p. putida polypeptide with an apparent molecular weight of 60,000 in sodium dodecyl sulfate-polyacrylamide gels. this polypeptide is presumed to be a methyl-accepting chemotaxis protein for several reasons: its molecular weight is similar to the molecular weights of escherichia coli methyl-acceptin ...	1989	2768186
degradation of phenol and m-toluate in pseudomonas sp. strain est1001 and its pseudomonas putida transconjugants is determined by a multiplasmid system.	the utilization of phenol, m-toluate, and salicylate (phe+, mtol+, and sal+ characters, respectively) in pseudomonas sp. strain est1001 is determined by the coordinated expression of genes placed in different plasmids, i.e., by a multiplasmid system. the natural multiplasmid strain est1001 is phenotypically unstable. in its phe-, mtol-, and sal- segregants, the plasmid dna underwent structural rearrangements without a marked loss of plasmid dna, and the majority of segregants gave revertants. th ...	1989	2768199
degradation of 2-bromo-, 2-chloro- and 2-fluorobenzoate by pseudomonas putida clb 250.	pseudomonas putida strain clb 250 (dsm 5232) utilized 2-bromo-, 2-chloro- and 2-fluorobenzoate as sole source of carbon and energy. degradation is suggested to be initiated by a dioxygenase liberating halide in the first catabolic step. after decarboxylation and rearomatization catechol is produced as a central metabolite which is degraded via the ortho-pathway. after inhibition of ring cleavage activities with 3-chlorocatechol, 2-chlorobenzoate was transformed to catechol in nearly stoichiometr ...	1989	2777062
identification of nucleotides critical for activity of the pseudomonas putida catbc promoter.	pseudomonas putida utilizes the catbc operon, which encodes cis,cis-muconate lactonizing enzyme i (mlei; ec 5.5.1.1) and muconolactone isomerase (mi; ec 5.3.3.4), for growth on benzoate as a sole carbon source. this operon is positively regulated, and the promoter is located 64 bp upstream of the catb translational start site. using site-specific mutagenesis, we identified nucleotides that influenced the induction of this promoter. promoter activity was monitored with the promoter probe vector p ...	1989	2779516
cloning, expression, and regulation of the pseudomonas cepacia protocatechuate 3,4-dioxygenase genes.	the genes for the alpha and beta subunits of the enzyme protocatechuate 3,4-dioxygenase (ec 1.13.11.3) were cloned from the pseudomonas cepacia dbo1 chromosome on a 9.5-kilobase-pair psti fragment into the broad-host-range cloning vector pro2317. the resultant clone was able to complement protocatechuate 3,4-dioxugenase mutations in p. cepacia, pseudomonas aeruginosa, and pseudomonas putida. expression studies showed that the genes were constitutively expressed and subject to catabolite repressi ...	1989	2808302
genetic organization and sequence of the pseudomonas cepacia genes for the alpha and beta subunits of protocatechuate 3,4-dioxygenase.	the locations of the genes for the alpha and beta subunits of protocatechuate 3,4-dioxygenase (ec 1.13.11.3) on a 9.5-kilobase-pair psti fragment cloned from the pseudomonas cepacia dbo1 chromosome were determined. this was accomplished through the construction of several subclones into the broad-host-range cloning vectors pro2317, pro2320, and pro2321. the ability of each subclone to complement mutations in protocatechuate 3,4-dioxygenase (pcaa) was tested in mutant strains derived from p. cepa ...	1989	2808303
[mutants of the plasmid for biodegradation of naphthalene, determining catechol oxidation via the meta-pathway].	most of the known naphthalene biodegradation plasmids determine the process of naphthalene degradation via salicylate and catechol using the meta pathway of catechol degradation. however, pseudomonas putida strains with plasmids pbs2, pbs216, pbs217 and npl-1 exert no activity of the enzymes involved in the meta pathway of catechol degradation. when 2-methylnaphthalene was added to the medium as a sole carbon source, mutants growing on this compound were isolated in the strains with the studied ...	1989	2811710
[preservation of the viability of opisthorchis eggs by joint cultivation with pseudomonas putida].	the effect of pseudomonas putida on opisthorchis' ova was studied with a view to assess the feasibility of using bacteria as biological agents against opisthorchiasis. experiments on mixed culture of the above-mentioned bacteria and helminths' ova demonstrated the lack of ovicidal effect of pseudomonas putida on the ova.	1989	2811755
autoradiographic determination of mass-transfer limitations in immobilized cell reactors.	pseudomonas putida cells were grown in confined volumes in dual-membrane immobilized cell reactors constructed from microporous polyethylene hollow fibers and silicone rubber tubules as a model system for the study of mass transport in microbial aggregates. local cell concentrations in the reactors reached 300 g dry mass/l. pulse-chase radioisotope labeling with (35)so(4) (2-) was used to estimate the rates of cell mass synthesis and degradation. sulfur incorporation consistently exceeded sulfur ...	1989	18588110
oxygen diffusivity in gel beads containing viable cells.	this article proposes a simple steady-state method for measuring the effective diffusion coefficient of oxygen (d(e)) in gel beads entrapping viable cells. we applied this method to the measurement of d(e) in ca- and ba-alginate gel beads entrapping saccharomyces cerevisiae and pseudomonas ovalis. the diffusivity of oxygen through gel beads containing viable cells was measured within an accuracy of +/-7% and found not to be influenced by cell density (0-30 g/l gel), cell type, and cell viability ...	1989	18588184
dynamics of continuous stirred-tank biochemical reactor utilizing inhibitory substrate.	the model of a continuous-stirred tank biochemical reactor was developed in which the instant uptake rate of substrate was used. the solutions of the model found for the oxidation of phenol by pseudomonas putida fitted the experimental data better than the results obtained from the models cited in the literature. the model enables control of the culture parameters so that the unwanted washout of the biomass from the bioreactor can be avoided. a review of the models cited in the literature is als ...	1988	18584592
uptake rate of phenol by pseudomonas putida grown in unsteady state.	the uptake rate of phenol by washed cells of pseudomonas putidagrown on phenol in fermenter in an un steady state, caused by the step increase of dilution rate and/or phenol concentration in the feed, was studied. the monod-haldane type equation was applied to fit the data and the best kinetic parameters were calculated by nonlinear least-square techniques. it was found that the minimum period of unsteady state required for induction of the phenol metabolic pathway was approximately 30 min. the ...	1988	18587828
radiometric determination of uranium accumulated in bacterial cells.	a method for the radiometric determination of uranium accumulated in bacterial cells was investigated. pseudomonas putida atcc 33015 was grown in a medium containing uranium in the form of uo(2)(no(3))(2). the cells were harvested by centrifugation, washed, resuspended in a buffer solution, ultrasonically disrupted and then suspended in unisolve-1. the radiation emitted by natural uranium isotopes (mainly (238)u and (234)u) was measured by liquid scintillation counting with natural uranium as an ...	1988	18964500
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 ...	1988	2971647
transcription of the fimbrial subunit gene and an associated transfer rna gene of pseudomonas aeruginosa.	gene fima encoding the structural subunit of the fimbriae of pseudomonas aeruginosa pak is located in the centre of a 1.2-kb hindiii genomic dna fragment [see also sastry et al., j. bacteriol. 164 (1985) 571-577], which in turn is located within a 6.2-kb ecori fragment. immediately downstream from fima is a putative threonine trna gene [dalrymple and mattick, biochem. int. 13 (1986) 547-553]. northern blotting experiments showed that fima is transcribed to an mrna of approx. 650 nucleotides, whi ...	1988	2452767
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 ...	1988	2830460
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 ...	1988	2831968
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 ...	1988	2832387
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 ...	1988	2832387
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 ...	1988	2833161
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 ...	1988	3046941
[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.	1988	3048271
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 ...	1988	3053681
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 ...	1988	3058208
[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 ...	1988	3058550
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 ...	1988	3063207
[nucleotide sequence of of rpob gene encoding the rna-polymerase beta-subunit in pseudomonas putida].		1988	3069413
[nucleotide sequence of the rpoc-gene coding for rna polymerase beta'-subunit of pseudomonas putida].		1988	3069416
cloning of genes for branched-chain keto acid dehydrogenase in pseudomonas putida.		1988	3071713
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 ...	1988	3073382
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 ...	1988	3075659
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 ...	1988	3076176
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 ...	1988	3076182
[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 ...	1988	3129337
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. ...	1988	3130804
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 ...	1988	3132906
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 ...	1988	3133958
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 ...	1988	3136164
[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 ...	1988	3137462
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 ...	1988	3139626
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.	1988	3141570
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 ...	1988	3144244
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.	1988	3146950
transcriptional regulation, nucleotide sequence, and localization of the promoter of the catbc operon in pseudomonas putida.	the catb and catc genes encode cis,cis-muconate lactonizing enzyme i (ec 5.5.1.1) and muconolactone isomerase (ec 5.3.3.4), respectively. these enzymes are required for the dissimilation of benzoate to beta-ketoadipate by pseudomonas putida and are under coordinate transcriptional regulation. by deletion analysis and the use of pkt240 as a promoter probe vector, we located a single promoter region for the catbc operon upstream of catb. rna-dna hybridization studies, together with reverse transcr ...	1988	2449420
genetic organization and transcriptional analysis of a major gene cluster involved in siderophore biosynthesis in pseudomonas putida wcs358.	in iron-limited environments, the plant-growth-stimulating pseudomonas putida wcs358 produces a yellow-green fluorescent siderophore called pseudobactin 358. the transcriptional organization and the iron-regulated expression of a major gene cluster involved in the biosynthesis and transport of pseudobactin 358 were analyzed in detail. the cluster comprises a region with a minimum length of 33.5 kilobases and contains at least five transcriptional units, of which some are relatively large. the di ...	1988	2450869
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 ...	1988	2838460
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 ...	1988	2841300
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 ...	1988	2841304
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 ...	1988	2842290
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 ...	1988	2842309
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 ...	1988	2843094
[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.	1988	2843421
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 ...	1988	2843518
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 ...	1988	2843820