Publications

purification and characterization of extracellular pectate lyase from bacillus subtilis.	bacillus subtilis strain so113 secretes a pectate lyase which is produced during the exponential death phase of growth, just before sporulation. this extracellular pectate lyase, which produces unsaturated products from polygalacturonate, was purified 35-fold from the culture supernatant of bacillus subtilis by a cm sephadex chromatography. it has an isoelectric point of about 9.6 and an mr of 42,000. optimum activity occurred at ph 8.4 and at 42 degrees c. calcium has a stimulative effect on th ...	1990	2126210
identification of plant-inducible genes in erwinia chrysanthemi 3937.	we present a method for identifying plant-inducible genes of erwinia chrysanthemi 3937. mutagenesis was done with the mu diipr3 transposon, which carries a promoterless neomycin phosphotransferase gene (npti), so upon insertion, the truncated gene can fuse to e. chrysanthemi promoters. mutants containing insertions in plant-inducible genes were selected for their sensitivity to kanamycin on minimal plates and for their acquired resistance to this antibiotic when an s. ionantha plant extract was ...	1990	2155205
characterization of transposon insertion out- mutants of erwinia carotovora subsp. carotovora defective in enzyme export and of a dna segment that complements out mutations in e. carotovora subsp. carotovora, e. carotovora subsp. atroseptica, and erwinia chrysanthemi.	soft-rotting erwinia spp. export degradative enzymes to the cell exterior (out+), a process contributing to their ability to macerate plant tissues. transposon (tn5, tn10, tn10-lacz) insertion out- mutants were obtained in erwinia carotovora subsp. carotovora 71 by using plasmid and bacteriophage lambda delivery systems. in these mutants, pectate lyases, polygalacturonase, and cellulase, which are normally excreted into the growth medium, accumulated in the periplasm. however, localization of th ...	1990	2160934
molecular cloning, nucleotide sequence, and marker exchange mutagenesis of the exo-poly-alpha-d-galacturonosidase-encoding pehx gene of erwinia chrysanthemi ec16.	the pehx gene encoding extracellular exo-poly-alpha-d-galacturonosidase (exopg; ec 3.2.1.82) was isolated from a genomic library of the pectate lyase-deficient erwinia chrysanthemi mutant um1005 (a nalr kanr delta pelabce derivative of ec16) by immunoscreening 2,800 escherichia coli hb101 transformants with an antibody against exopg protein. the cloned pehx gene was expressed highly from its own promoter in e. coli, and most of the enzyme was localized in the periplasm. the nucleotide sequence o ...	1990	2168372
protease secretion by erwinia chrysanthemi: the specific secretion functions are analogous to those of escherichia coli alpha-haemolysin.	a 5.5 kb dna fragment carrying the functions necessary for the specific secretion of the extracellular metalloproteases b and c produced by the gram-negative phytopathogenic bacterium erwinia chrysanthemi has been sequenced. the fragment contains four transcribed and translated genes: inh, which codes for a protease inhibitor and is not required for protease secretion, and prtd, prte and prtf, which share significant homology with the hlyb, hlyd and tolc genes required for alpha-haemolysin secre ...	1990	2184029
molecular cloning and characterization of an erwinia carotovora subsp. carotovora pectin lyase gene that responds to dna-damaging agents.	reca-mediated production of pectin lyase (pnl) and the bacteriocin carotovoricin occurs in erwinia carotovora subsp. carotovora 71 when this organism is subjected to agents that damage or inhibit the synthesis of dna. the structural gene pnla was isolated from a strain 71 cosmid gene library following mobilization of the cosmids into a moderate pnl producer, strain 193. the cosmid complemented pnl::tn5 but not ctv::tn5 mutations. a constitutive level of pnl activity was detected in reca+ and rec ...	1990	2188956
preliminary crystallographic analysis of the plant pathogenic factor, pectate lyase c from erwinia chrysanthemi.	pectate lyases are saccharide-binding enzymes that degrade plant cell walls. one pectate lyase from erwinia chrysanthemi (ec16), termed pectate lyase c, has been crystallized from ammonium sulfate. the preliminary x-ray diffraction analysis indicates that the crystals belong to the orthorhombic space group p2(1)2(1)2(1), with unit cell dimensions, a = 73.4 a, b = 80.3 a, and c = 95.1 a. the crystals diffract to a resolution of 2.2 a and have one molecule/asymmetric unit.	1990	2195018
[fusion of erwinia chrysanthemi spheroplasts in an electric fields].	a new approach has been elaborated for electrofusion of erwinia chrysanthemi spheroplasts. the new approach consists of superimposition of high voltage impulses on the pellet of tightly contacting cells in the course of centrifugation. the mixture of spheroplasts of two genetically marked strains was placed into the special centrifuge chambers and spinned for 15 min at 2500 g to get a compressed pellet between chamber electrodes. three successive pulses of 6.6 kv/cm amplitude and 30 microseconds ...	1990	2199828
cloning of genes encoding extracellular metalloproteases from erwinia chrysanthemi ec16.	a 14-kilobase bamhi-ecori dna fragment cloned from erwinia chrysanthemi ec16 contained a gene encoding a metalloprotease inhibitor as well as three tandem prt genes encoding metalloproteases. the prt genes were separated from the inhibitor gene by a ca. 4-kilobase region that was necessary for extracellular secretion of the proteases. when individually subcloned downstream from vector promoters, the three prt genes each led to substantial extracellular secretion of the proteases by escherichia c ...	1990	2211513
protein secretion in gram-negative bacteria. the extracellular metalloprotease b from erwinia chrysanthemi contains a c-terminal secretion signal analogous to that of escherichia coli alpha-hemolysin.	the secretion signal of extracellular metalloprotease b that is secreted without a signal peptide by the gram-negative phytopathogenic bacterium erwinia chrysanthemi is shown by deletion and gene fusion analyses to be located within the last 40 c-terminal amino acids. secretion of a peptide containing only this region of the protease requires the same three secretion factors (prtd, prte, and prtf) that were previously shown to be required for the secretion of the full-length protease. this secre ...	1990	2211614
analysis of the erwinia chrysanthemi arb genes, which mediate metabolism of aromatic beta-glucosides.	erwinia chrysanthemi is one of the few members of the family enterobacteriaceae that is capable of metabolizing most of the naturally occurring beta-glucosides. we previously isolated the clb genes, which allow the use of the disaccharide cellobiose as well as the aromatic beta-glucosides arbutin and salicin. we report here the isolation of the arb genes, which permit fermentation of the aromatic beta-glucosides only. establishment of a functional arb system in escherichia coli depended on the p ...	1990	2228958
non-randomised study comparing toxicity of escherichia coli and erwinia asparaginase in children with leukaemia.	seven hundred fifty-eight unselected children entered into the united kingdom medical research council acute lymphoblastic leukaemia ukall viii study and trial were studied for differences in early treatment-related toxicity according to the type of intramuscular l-asparaginase received. two hundred seventy-five received a product obtained from escherichia coli and 483 the enzyme from erwinia chrysanthemi. the e. coli patients had a significantly higher incidence of neurotoxicity, pancreatitis, ...	1990	2233523
molecular cloning of the structural gene for exopolygalacturonate lyase from erwinia chrysanthemi ec16 and characterization of the enzyme product.	the ability of erwinia chrysanthemi to cause soft-rot diseases involving tissue maceration in many plants has been linked to the production of endo-pectate lyase e. chrysanthemi ec16 mutant um1005, however, contains deletions in the pel genes that encode the known endopectate lyases, yet still macerates plant tissues. in an attempt to identify the remaining macerating factor(s), a gene library of um1005 was constructed in escherichia coli and screened for pectolytic activity. a clone (ppnl5) was ...	1990	2254266
serodiagnosis of helicobacter pylori infection in childhood.	sera from 100 children (ages, 6 to 16 years) presenting with upper gastrointestinal symptoms were examined for antibodies to helicobacter pylori by enzyme-linked immunosorbent assay (elisa) based on crude, loosely cell-associated antigens and a partially purified urease antigen preparation. all children underwent endoscopy, and 20 children were shown to have h. pylori infection by histology or direct culture. serum anti-h. pylori immunoglobulin g (igg) levels (crude antigen) were clearly raised ...	1990	2279995
the virulence-associated chrysobactin iron uptake system of erwinia chrysanthemi 3937 involves an operon encoding transport and biosynthetic functions.	the iron assimilation system of erwinia chrysanthemi 3937 is mediated by the catechol-type siderophore chrysobactin and the outer membrane transport protein fct. we generated a variety of subclones in high- and low-copy-number vectors from a wild-type recombinant cosmid shown previously to carry the gene cluster fct-cbsa, cbsb, cbsc, cbse encoding chrysobactin transport and biosynthetic functions, respectively. we studied their expression in escherichia coli enterobactin-deficient enta, entb, en ...	1991	1657869
conjugational transfer of recombinant dna in cultures and in soils: host range of pseudomonas putida tol plasmids.	recombinant tol plasmid pwwo-eb62 allows pseudomonas putida to grow on p-ethylbenzoate. this plasmid can be transferred to other microorganisms, and its catabolic functions for the metabolism of alkylbenzoates are expressed in a limited number of gram-negative bacteria, including members of pseudomonad rrna group i and escherichia coli. transfer of the recombinant plasmid to erwinia chrysanthemi was observed, but transconjugants failed to grow on alkylbenzoates because they lost catabolic functi ...	1991	1660698
simultaneous expression of a bacteriophage mu transposase and repressor: a way of preventing killing due to mini-mu replication.	in vitro studies of bacteriophage mu transposition have shown that the phage-encoded transposase and repressor bind the same sequences on the phage genome. we attempted to test that prediction in vivo and found that mu repressor directly inhibits transposition. we also found that, in the absence of repressor, constitutive expression of mu transposition functions pa and pb is lethal in escherichia coli strains lysogenic for a mini-mu and that this is the result of intensive replication of the min ...	1991	1662754
cellulase egz of erwinia chrysanthemi: structural organization and importance of his98 and glu133 residues for catalysis.	biochemical, genetic and primary sequence analyses of the erwinia chrysanthemi endoglucanase egz allowed us to identify two functional domains and to locate their boundaries. the catalytic domain extends from residue 1 to 288, while a domain required for egz to bind to microcrystalline cellulose lies from residues 324 to 385. each domain was found capable of functioning in the absence of the other. a region rich in pro, thr, and ser residues links both domains and appeared to be susceptible to p ...	1991	1677466
analysis of an erwinia chrysanthemi gene cluster involved in pectin degradation.	a group of four genes of erwinia chrysanthemi involved in pectin degradation has been characterized. these four genes form independent transcription units and are regulated by the negative regulatory gene, kdgr. the functions of two of these genes are known: kdud codes for the 2-keto-3-deoxygluconate oxydoreductase and kdul for the 5-keto-4-deoxyuronate isomerase, two enzymes of the pectin degradation pathway. kdgc has 36% homology with pectate lyase genes of the periplasmic family but its produ ...	1991	1766386
genetic analysis of the erwinia chrysanthemi 3937 chrysobactin iron-transport system: characterization of a gene cluster involved in uptake and biosynthetic pathways.	twenty of the twenty-two mudii1734 insertions impairing the chrysobactin iron-assimilation system of erwinia chrysanthemi 3937 were localized to a 50 kbp genomic insert contained in the r-prime plasmid, r'4 (enard et al., 1988). using the conjugative plasmid pulb110 (rp4::mini-mu) and the generalized transducing phage phi ec2, we located this iron-transport region and the two unlinked mutations on the chromosome linkage map. chrysobactin is a catechol-type siderophore and, as we have previously ...	1991	1787788
[temperate sm phage from pseudomonas aeruginosa as a vector for cloning genetic information].	the recombinant bacteriophages with the genomes containing the dna fragments of bacteria erwinia chrysanthemi, including the pectatelyase gene, were constructed on the base of pseudomonas aeruginosa temperate bacteriophage sm. the gene transferred into pseudomonas aeruginosa pao1 cells by transfection is expressed in the new bacterial host. the restriction maps of the recombinant bacteriophages are constructed and the position of an insert is defined. bacteriophage sm was found to be capable of ...	1991	1787841
characterization, localization and transmembrane organization of the three proteins prtd, prte and prtf necessary for protease secretion by the gram-negative bacterium erwinia chrysanthemi.	erwinia chrysanthemi, a gram-negative phythopathogenic bacterium, secretes two related extracellular metalloproteases, b and c, which do not have n-terminal signal sequences. the specific pathway by which they are secreted, which has been reconstituted in escherichia coli, comprises three proteins -- prtd, prte and prtf. hybrid proteins containing segments of these proteins fused to the c-terminus of protease b were purified and used to immunize rabbits. the antisera thus obtained were used to s ...	1991	1791757
extracellular secretion of pectate lyase by the erwinia chrysanthemi out pathway is dependent upon sec-mediated export across the inner membrane.	the plant pathogenic enterobacterium erwinia chrysanthemi ec16 secretes several extracellular, plant cell wall-degrading enzymes, including pectate lyase isozyme pele. secretion kinetics of 35s-labeled pele indicated that the precursor of pele was rapidly processed by the removal of the amino-terminal signal peptide and that the resulting mature pele remained cell bound for less than 60 s before being secreted to the bacterial medium. pele-phoa (alkaline phosphatase) hybrid proteins generated in ...	1991	1829728
characterization of kdgr, a gene of erwinia chrysanthemi that regulates pectin degradation.	erwinia chrysanthemi is a phytopathogenic enterobacterium able to degrade the pectic fraction of plant cell walls. the kdgr negative regulatory gene controls all the genes involved in pectin catabolism, including the pel genes encoding pectate lyases. the e. chrysanthemi kdgr regulatory gene was subcloned in escherichia coli where it was shown to be functional, since it repressed the expression of a pele::uida fusion. the nucleotide sequence of kdgr contained an open reading frame of 918bp prece ...	1991	1840643
a gene showing sequence similarity to pectin esterase is specifically expressed in developing pollen of brassica napus. sequences in its 5' flanking region are conserved in other pollen-specific promoters.	differential screening of a brassica napus genomic library led to the isolation of the clone named bp 19 containing a gene which is highly expressed during microspore development. the accumulation of bp19 mrna starts in uninucleate microspores, increases during development reaching a peak in the late stages but declines considerably in mature pollen. the nucleotide sequence of the entire coding region and of extended portions of the 5' and 3' flanking regions was determined. several homologous c ...	1991	1868195
inducing properties of analogs of 2-keto-3-deoxygluconate on the expression of pectinase genes of erwinia chrysanthemi.	in erwinia chrysanthemi, all the genes involved in pectin degradation are controlled by the negative regulatory gene kdgr. 2-keto-3-deoxy-gluconate (kdg) is the inducing molecule that interacts with kdgr to allow the expression of all the genes of the kdg regulon. the inducing properties on the expression of genes regulated by kdgr of various analogs and derivatives of kdg were tested. all the inducers share the common moiety cooh-co-ch2-choh-c-c included in a pyranic cycle. our results show tha ...	1991	1874406
the secretion genes of pseudomonas aeruginosa alkaline protease are functionally related to those of erwinia chrysanthemi proteases and escherichia coli alpha-haemolysin.	the extracellular alkaline protease produced by pseudomonas aeruginosa is secreted by a specific pathway, independent of the pathway used by most of the other extracellular proteins of this organism. secretion of this protease is dependent on the presence of several genes located adjacent to the apr gene. complementation studies have shown that prtd, e, and f, the three secretion functions for erwinia chrysanthemi proteases b and c (létoffé et al., 1990), can mediate the secretion of the alkalin ...	1991	1904127
erwinia carotovora subsp. carotovora extracellular protease: characterization and nucleotide sequence of the gene.	the prt1 gene encoding extracellular protease from erwinia carotovora subsp. carotovora ec14 in cosmid pca7 was subcloned to create plasmid psk1. the partial nucleotide sequence of the insert in psk1 (1,878 bp) revealed a 1,041-bp open reading frame (orf1) that correlated with protease activity in deletion mutants. orf1 encodes a polypeptide of 347 amino acids with a calculated molecular mass of 38,826 da. escherichia coli transformed with psk1 or psk23, a subclone of psk1, produces a protease ( ...	1991	1917878
sequence analysis of the cellulase-encoding cely gene of erwinia chrysanthemi: a possible case of interspecies gene transfer.	the erwinia chrysanthemi (strain 3937) cely gene encoding the minor endoglucanase (egy) was sequenced. the analysis of the upstream region allowed us to identify an in vivo active promoter recognized by the ntra (sigma 54) holoenzyme. no similarity was found between the predicted amino acid (aa) sequences of egy and either the er. chrysanthemi major endoglucanase, egz, or the er. carotovora cels endoglucanase. in contrast, a very high level of identity, both at the nucleotide and the predicted a ...	1991	1937031
cloned erwinia chrysanthemi out genes enable escherichia coli to selectively secrete a diverse family of heterologous proteins to its milieu.	the out genes of the enterobacterial plant pathogen erwinia chrysanthemi are responsible for the efficient extracellular secretion of multiple plant cell wall-degrading enzymes, including four isozymes of pectate lyase, exo-poly-alpha-d-galacturonosidase, pectin methylesterase, and cellulase. out- mutants of er. chrysanthemi are unable to export any of these proteins beyond the periplasm and are severely reduced in virulence. we have cloned out genes from er. chrysanthemi in the stable, low-copy ...	1991	1992458
cloning and expression in escherichia coli of the serratia marcescens metalloprotease gene: secretion of the protease from e. coli in the presence of the erwinia chrysanthemi protease secretion functions.	the serratia marcescens extracellular protease sm is secreted by a signal peptide-independent pathway. when the prtsm gene was cloned and expressed in escherichia coli, the cells did not secrete protease sm. the lack of secretion could be very efficiently complemented by the erwinia chrysanthemi protease b secretion apparatus constituted by the prtd, prte, and prtf proteins. as with protease b and alpha-hemolysin, the secretion signal was located within the last 80 amino acids of the protease. t ...	1991	2007544
molecular cloning and sequencing of a pectinesterase gene from pseudomonas solanacearum.	two pectinesterase-positive escherichia coli clones, differing in expression levels, were isolated from a genomic library of pseudomonas solanacearum. both clones contained a common dna fragment which included the pectinesterase-encoding region. the different expression levels found with the two clones could be ascribed to different positioning of the pectinesterase gene with respect to a vector promoter. restriction analysis, subcloning, and further exonuclease deletion mapping revealed that th ...	1991	2045776
production of pectin methylesterase from erwinia chrysanthemi b374 in bacillus subtilis.	the gene coding for pectin methylesterase (pme) of erwinia chrysanthemi b374 (pme) was cloned by a polymerase chain reaction. the pme gene was expressed in bacillus subtilis using a secretion vector based on the promoter and signal sequence of the alpha-amylase gene from b. amyloliquefaciens. the cultivation of b. subtilis cells carrying the cloned pme resulted in efficient secretion of pme into the culture medium based on enzymatic and sodium dodecyl sulphate-poly-acrylamide gel electrophoresis ...	1991	1367534
rapid large-scale preparation of recombinant erwinia chrysanthemi l-asparaginase.	l-asparaginase from erwinia provides an alternative to the enzyme from e. coli for the effective treatment of acute lymphoblastic leukaemia. a procedure was required for the large-scale partial purification of the recombinant erwinia enzyme cloned and expressed in erwinia. enzyme was extracted from erwinia at high ph and extraneous protein precipitated at low ph. s-sepharose ff was selected as the medium of choice for the chromatography step since it was adequate for the high flow rates required ...	1992	1368993
iron(iii) complexes of chrysobactin, the siderophore of erwinia chrysanthemi.	the phytopathogenic bacterium erwinia chrysanthemi produces the monocatecholate siderophore chrysobactin under conditions of iron deprivation. only the catecholate hydroxyl groups participate in metal coordination, and chrysobactin is therefore unable to provide full 1:1 coordination of fe(iii). the stoichiometry in aqueous solution is a variable dependent on ph and metal/ligand ratio, in addition to being concentration dependent. at neutral ph and concentrations of about 0.1 mm, ferric chrysoba ...	1992	1392469
analysis of the regulation of the pelbc genes in erwinia chrysanthemi 3937.	erwinia chrysanthemi secretes five major isoenzymes of pectate lyases encoded by the pelabcde genes. the nucleotide sequence of the region surrounding the pelb gene of e. chrysanthemi 3937 was determined, including the regulatory regions involved in pelb and pelc expression. analysis of the transcripts showed that transcription of pelb or pelc gave, in both cases, only one transcript. the transcription initiation sites of both pelb and pelc were precisely determined as well as the position of th ...	1992	1406275
synthesis and secretion of an erwinia chrysanthemi pectate lyase in saccharomyces cerevisiae regulated by different combinations of bacterial and yeast promoter and signal sequences.	nine different expression-secretion cassettes, comprising novel combinations of yeast and bacterial gene promoters and secretion signal sequences, were constructed and evaluated. a pectate lyase-encoding gene (pele) from erwinia chrysanthemi was inserted between each one of these expression-secretion cassettes and a yeast gene terminator, generating recombinant yeast-integrating shuttle plasmids pams1 through pams9. these yip5-derived plasmids were transformed and stably integrated into the geno ...	1992	1427097
sequence of a cluster of genes controlling synthesis and secretion of alkaline protease in pseudomonas aeruginosa: relationships to other secretory pathways.	a genetic locus implicated in the synthesis and secretion of alkaline protease (apr) in pseudomonas aeruginosa has been previously described [guzzo et al., j. bacteriol. 172 (1990) 942-948]. the nucleotide sequence of the dna fragment encoding these functions was determined and revealed the existence of five open reading frames: apra, the structural gene encoding apr; apri, which encodes a protease inhibitor; and aprd, apre, aprf whose products are involved in protease secretion. the aprd, apre ...	1992	1427098
analysis of eight out genes in a cluster required for pectic enzyme secretion by erwinia chrysanthemi: sequence comparison with secretion genes from other gram-negative bacteria.	many extracellular proteins produced by erwinia chrysanthemi require the out gene products for transport across the outer membrane. in a previous report (s. y. he, m. lindeberg, a. k. chatterjee, and a. collmer, proc. natl. acad. sci. usa 88:1079-1083, 1991) cosmid pcpp2006, sufficient for secretion of erwinia chrysanthemi extracellular proteins by escherichia coli, was partially sequenced, revealing four out genes sharing high homology with pulh through pulk from klebsiella oxytoca. the nucleot ...	1992	1429461
environmental conditions affect transcription of the pectinase genes of erwinia chrysanthemi 3937.	to depolymerize plant pectin, the phytopathogenic enterobacterium erwinia chrysanthemi produces a series of enzymes which include a pectin-methyl-esterase encoded by the pem gene and five isoenzymes of pectate lyases encoded by the five genes pela, pelb, pelc, peld, and pele. we have constructed transcriptional fusions between the pectinase gene promoters and the uida gene, encoding beta-glucuronidase, to study the regulation of these e. chrysanthemi pectinase genes individually. the transcripti ...	1992	1447147
some of the out genes involved in the secretion of pectate lyases in erwinia chrysanthemi are regulated by kdgr.	the out genes of erwinia chrysanthemi are required for the translocation across the outer membrane of pectate lyases and cellulases. we present the characterization and the nucleotide sequence of five genes of the out cluster. the products of outs, b, c, d and e have significant homology with the puls, b, c, d and e proteins necessary to the secretion of pullulanase in klebsiella pneumoniae. an open reading frame, outt, located between outb and outc has no homology with the pul cluster but is in ...	1992	1453958
cloning, nucleotide sequence and characterization of the gene encoding the erwinia chrysanthemi b374 prta metalloprotease: a third metalloprotease secreted via a c-terminal secretion signal.	erwinia chrysanthemi, a phytopathogenic enterobacterium, secretes three proteases (prta, prtb and prtc) into the extracellular medium. the gene encoding the 50 kda protease, prta, was subcloned from a recombinant cosmid carrying a fragment of the e. chrysanthemi b374 chromosome. prta was shown to be located immediately 3' to the structural genes for the other two extracellular proteases. the amino acid sequence of prta, as predicted from the prta nucleotide sequence, showed a high level of homol ...	1992	1494344
negative transcriptional control of iron transport in erwinia chrysanthemi involves an iron-responsive two-factor system.	systemic virulence of the phytopathogen erwinia chrysanthemi 3937 requires a functional iron assimilation system which, in this enterobacterium, is mediated by the siderophore chrysobactin and the outer membrane transport protein fct. we investigated the regulation of this system by iron. no direct similarity with the escherichia coli fur gene was found. insertional mutagenesis allowed isolation of a regulatory mutant which expressed chrysobactin and two other high-affinity iron transport system ...	1992	1508046
cloning and characterization of a phospholipase gene from erwinia chrysanthemi ec16.	a single gene (plca) was cloned from a cosmid library of erwinia chrysanthemi ec16 dna that encoded an extracellular phospholipase. the gene was subcloned and dna sequence data showed the presence of a single open reading frame encoding a protein with a predicted size of 39 kda. the coding region was g+c-rich and the protein had a predicted basic isoelectric point. the protein showed no significant homology with others in the pir library, including other phospholipases. when overexpressed in esc ...	1992	1545703
secretion of the rhizobium leguminosarum nodulation protein nodo by haemolysin-type systems.	the rhizobium leguminosarum biovar viciae nodulation protein nodo is partially homologous to haemolysin of escherichia coli and, like haemolysin, is secreted into the growth medium. the nodo protein can be secreted by a strain of e. coli carrying the cloned nodo gene plus the haemolysin secretion genes hlybd, in a process that also requires the outer membrane protein encoded by tolc. the related protease secretion genes, prtdef, from erwinia chrysanthemi also enable e. coli to secrete nodo. the ...	1992	1545707
purification and functional characterization of the kdgr protein, a major repressor of pectinolysis genes of erwinia chrysanthemi.	the phytopathogenicity of the enterobacterium erwinia chrysanthemi chiefly results from its capacity to degrade pectin, which is the major component of plant cell walls. this degradation requires the product of 12 genes which constitute independent transcriptional units. all these genes, including kdgt which encodes the 2-keto-3-deoxygluconate (kdg) transport system, are negatively regulated by the kdgr protein. the e. chrysanthemi kdgr gene was cloned into an expression vector and overexpressed ...	1992	1545709
differential depolymerization mechanisms of pectate lyases secreted by erwinia chrysanthemi ec16.	the four pectate lyases (ec 4.2.2.2) secreted by erwinia chrysanthemi ec16 have been individually produced as recombinant enzymes in escherichia coli. oligogalacturonates formed from polygalacturonic acid during reactions catalyzed by each enzyme have been determined by high-performance liquid chromatography analysis. pla catalyzes the formation of a series of oligomers ranging from dimer to dodecamer through a random endolytic depolarization mechanism. plb and plc are trimer- and tetramer-gener ...	1992	1548242
production of active serratia marcescens metalloprotease from escherichia coli by alpha-hemolysin hlyb and hlyd.	serratia marcescens produces an abundant extracellular metalloprotease. the gene for this protease had previously been cloned and expressed in escherichia coli, in which no functional protease could be found. however, the protease gene carries the lxggxgnd repeat motif found in alpha-hemolysin and other proteins secreted by homologous systems. using a dual-plasmid complementation system, we show that the alpha-hemolysin hlyb and hlyd transport determinants are sufficient to allow secretion and a ...	1992	1551853
stereochemistry of the hydrolysis reaction catalyzed by endoglucanase z from erwinia chrysanthemi.	endoglucanase z from the phytopathogenic bacterium erwinia chrysanthemi (strain 3937) was purified by affinity chromatography on microcrystalline cellulose avicel ph101. a kinetic characterization using p-nitrophenyl beta-d-cellobioside and p-nitrophenyl beta-d-lactosde as substrates was conducted: endoglucanase z exhibited km values of 3 mm and 7.5 mm and vm values of 129 and 40 nmol.min-1.mg-1 towards p-nitrophenyl beta-d-cellobioside (kcat = 0.1 s-1) and p-nitrophenyl beta-d-lactoside (kcat = ...	1992	1563515
purification of the acidic pectate lyase and nucleotide sequence of the corresponding gene (pela) of erwinia chrysanthemi strain 3937.	the pela gene from erwinia chrysanthemi strain 3937, which encodes the acidic pectate lyase, pla, has been sequenced and characterized. the structural gene consists of a 1179 bp open reading frame encoding a polypeptide of 41,555 da, which includes an n-terminal signal peptide. the deduced amino acid sequence shows a protein very similar to some pls already sequenced. cloning of the pela gene behind the lacz promoter of the vector ptz19r allowed overexpression of pla into a derivative of strain ...	1992	1593262
rapid detection of members of the family enterobacteriaceae by a monoclonal antibody.	six monoclonal antibodies directed against enterobacteria were produced and characterized. the specificity of one of these antibodies (cx9/15; immunoglobulin g2a) was studied by indirect immunofluorescence against 259 enterobacterial strains and 125 other gram-negative bacteria. all of the enterobacteria were specifically recognized, the only exception being erwinia chrysanthemi (one strain tested). bacteria not belonging to members of the family enterobacteriaceae were not detected, except for ...	1992	1622220
ferric iron uptake in erwinia chrysanthemi mediated by chrysobactin and related catechol-type compounds.	erwinia chrysanthemi 3937 possesses a saturable, high-affinity transport system for the ferric complex of its native siderophore chrysobactin, [n-alpha-(2,3-dihydroxybenzoyl)-d-lysyl-l-serine]. uptake of 55fe-labeled chrysobactin was completely inhibited by respiratory poison or low temperature and was significantly reduced in rich medium. the kinetics of chrysobactin-mediated iron transport were determined to have apparent km and vmax values of about 30 nm and of 90 pmol/mg.min, respectively. i ...	1992	1624465
secretion of cyaa-prtb and hlya-prtb fusion proteins in escherichia coli: involvement of the glycine-rich repeat domain of erwinia chrysanthemi protease b.	protease b from erwinia chrysanthemi was shown previously to have a c-terminal secretion signal located downstream of a domain that contains six glycine-rich repeats. this domain is conserved in all known bacterial proteins secreted by the signal peptide-independent pathway. the role of these repeats in the secretion process is controversial. we compared the secretion processes of various heterologous polypeptides fused either directly to the signal or separated from it by the glycine-rich domai ...	1992	1629152
distribution of an l-isoaspartyl protein methyltransferase in eubacteria.	a protein carboxyl methyltransferase (ec 2.1.1.77) that recognizes age-damaged proteins for potential repair or degradation reactions has been found in all vertebrate tissues and cells examined to date. this enzyme catalyzes the transfer of methyl groups from s-adenosylmethionine to the carboxyl groups of d-aspartyl or l-isoaspartyl residues that are formed spontaneously from normal l-aspartyl and l-asparaginyl residues. a similar methyltransferase has been found in two bacterial species, escher ...	1992	1729230
nucleotide sequences of the arb genes, which control beta-glucoside utilization in erwinia chrysanthemi: comparison with the escherichia coli bgl operon and evidence for a new beta-glycohydrolase family including enzymes from eubacteria, archeabacteria, and humans.	the phytopathogenic bacterium erwinia chrysanthemi, unlike other members of the family enterobacteriaceae, is able to metabolize the beta-glucosides, arbutin, and salicin. a previous genetic analysis of the e. chrysanthemi arb genes, which mediate beta-glucoside metabolism, suggested that they were homologous to the escherichia coli k-12 bgl genes. we have now determined the nucleotide sequence of a 5,065-bp dna fragment containing three genes, arbg, arbf, and arbb. deletion analysis, expression ...	1992	1732212
a fourth metalloprotease gene in erwinia chrysanthemi.	erwinia chrysanthemi, a gram-negative phytopathogenic bacterium, was previously shown to secrete 3 related extracellular metalloproteases, a, b and c via a specific signal-peptide-independent pathway. a new gene (prtg) encoding a fourth, 52-kda metalloprotease was identified on the same recombinant cosmid (pew1) that carries the genes for the previously described proteases (prta, prtb and prtc), for the specific secretion factors (prtd, prte and prtf) and for a protease inhibitor (inh) cloned fr ...	1992	1299839
analysis of the pele promoter in erwinia chrysanthemi ec16.	the pele gene of erwinia chrysanthemi strain ec16 encodes an extracellular pectate lyase protein that is important in virulence on plants. control of pele expression is complex, because the gene is regulated by catabolite repression, substrate induction, and growth-phase inhibition. a tn7-lux reporter gene system was employed to define dna sequences comprising the pele promoter. when ec16 cells were grown on medium containing sodium polypectate, pele transcriptional start sites were observed onl ...	1992	1319773
construction of a tn7-lux system for gene expression studies in gram-negative bacteria.	a tn7-lux system was developed for gene expression studies in gram- bacteria. the plasmids constructed, phsk728 and phsk729, have the following features: (1) a promoterless vibrio fischeri lux operon as a reporter system; (2) multiple cloning sites (mcs) ahead of the lux operon, in opposite orientation for the cloning of promoter fragments; (3) a transcriptional terminator ahead of the mcs and translational stop codons in all reading frames before the translational start of the luxc gene; (4) a ...	1992	1333438
iron deficiency induced by chrysobactin in saintpaulia leaves inoculated with erwinia chrysanthemi.	in this communication, we examine the fate of iron during soft rot pathogenesis caused by erwinia chrysanthemi on its host, saintpaulia ionantha. the spread of soft rot caused by this enterobacterium was previously shown to depend on a functional genetic locus encoding a high-affinity iron assimilation system involving the catechol-type siderophore chrysobactin. leaf intercellular fluid from healthy plants was analyzed with regard to the iron content and its availability for bacterial growth. it ...	1993	12231882
characterization of the prta and prtb genes of erwinia chrysanthemi ec16.	two tandem metalloprotease-encoding structural genes, prta and prtb, were sequenced from erwinia chrysanthemi ec16. these were highly homologous to previously reported genes from the same bacteria, as well as to three other metalloprotease-encoding genes from enteric bacteria. the three tandem prt structural genes from strain ec16 were closely linked to a cluster of genes previously found to be essential for extracellular secretion of the metalloproteases.	1993	8224883
identification of two components of the serratia marcescens metalloprotease transporter: protease sm secretion in escherichia coli is tolc dependent.	the serratia marcescens metalloprotease (protease sm) belongs to a family of proteins secreted from gram-negative bacteria by a signal peptide-independent pathway which requires a specific transporter consisting of three proteins: two in the inner membrane and one in the outer membrane. the prtdsm and prtesm genes encoding the two s. marcescens inner membrane components were cloned and expressed in escherichia coli. their nucleotide sequence revealed high overall homology with the two analogous ...	1993	8226679
molecular analysis of the major cellulase (celv) of erwinia carotovora: evidence for an evolutionary "mix-and-match" of enzyme domains.	the structural gene for the major cellulase of erwinia carotovora subspecies carotovora (ecc) was isolated and expressed in escherichia coli. sequencing of the gene (celv) revealed a typical signal sequence and two functional domains in the enzyme; a catalytic domain linked by a short proline/threonine-rich linker to a cellulose-binding domain (cbd). the deduced amino acid sequence of the catalytic domain showed homology with cellulases of family a, including enzymes from bacillus spp. and erwin ...	1993	8246888
pectate lyase from bacillus subtilis: molecular characterization of the gene, and properties of the cloned enzyme.	pectate lyases (pl) initiate soft-rot diseases in plants by cleaving pectin which is the major component of the plant cell wall. the present paper reports the first cloning and characterization of a pectate lyase (pel) gene from the bacillus genus. this gene was isolated from a bacillus subtilis genomic library constructed in puc18 as vector and escherichia coli as host. by southern hybridization this gene was shown to be present in a single copy in the b. subtilis genome. the nucleotide sequenc ...	1993	8262178
abc transporters: bacterial exporters.	the abc transporters (also called traffic atpases) make up a large superfamily of proteins which share a common function and a common atp-binding domain. abc transporters are classified into three major groups: bacterial importers (the periplasmic permeases), eukaryotic transporters, and bacterial exporters. we present a comprehensive review of the bacterial abc exporter group, which currently includes over 40 systems. the bacterial abc exporter systems are functionally subdivided on the basis o ...	1993	8302219
uptake of galacturonic acid in erwinia chrysanthemi ec16.	uptake of [14c]galacturonic acid in erwinia chrysanthemi was found to be stimulated during growth on pectin and its degradation products, saturated digalacturonic acid and galacturonic acid. cells isolated from macerated potato tissue also showed increased levels of uptake activity for this molecule compared with those showed by glycerol-grown cells. uptake was found to be an active process, and it displayed saturation kinetics. an escherichia coli galacturonic acid transport mutant harboring th ...	1993	8320243
molecular cloning and characterization of 13 out genes from erwinia carotovora subspecies carotovora: genes encoding members of a general secretion pathway (gsp) widespread in gram-negative bacteria.	the chemical mutagen ethylmethanesulphonate (ems) has been used to generate mutants of erwinia carotovora subspecies carotovora which are defective in the secretion of pectinases (pel) and cellulases (cel) but unaltered for protease (prt) secretion. such mutants, called out-, still synthesize pel and cel but these enzymes accumulate within the periplasm. cosmid clones carrying wild-type e. carotovora ssp. carotovora dna, identified by their ability to restore the out+ phenotype when transferred ...	1993	8326859
characterization of the nucm gene coding for a nuclease of the phytopathogenic bacteria erwinia chrysanthemi.	the gene nucm encoding a nuclease was cloned from a genomic library of erwinia chrysanthemi. the nucm gene was subcloned, and mutagenized by insertion of a uida-kanr cartridge. this mutation was introduced by recombination into the erwinia chrysanthemi chromosome. the nucm mutant lost nucm activity when tested on a dna plate after 24 hours, but still possessed secondary weak nuclease activity. the nucleotide sequence of nucm was determined. it presents a 798 bp open reading frame, coding for a 2 ...	1993	8332061
a left-handed crossover involved in amidohydrolase catalysis. crystal structure of erwinia chrysanthemi l-asparaginase with bound l-aspartate.	the crystal structure of l-asparaginase from erwinia chrysanthemi in the presence and absence of l-aspartate was determined at 1.8 a resolution. conserved residues in a left-handed crossover (a rare occurrence in protein structures) link pairs of dimers into the catalytically active tetrameric form of the enzyme. the structure of era containing bound aspartic acid shows that this unusual strand connectivity is an essential part of the active site architecture, responsible for releasing the produ ...	1993	8348975
structure of an extracellular polysaccharide produced by erwinia chrysanthemi.	erwinia chrysanthemi pv zeae strain sr260, a phytopathogen of corn, produced from lactose an acidic extracellular polysaccharide which was purified and found to consist of l-rhamnose, d-mannose, d-glucose, and d-glucuronic acid in the ratio of 3:1:1:1. a combination of chemical (carboxyl-group reduction, methylation analysis, periodate oxidation, smith degradation, and lithium-ethylenediamine degradation) and physical (1 and 2d nmr spectroscopy) methods revealed that the polysaccharide is compos ...	1993	8370026
characterization and overexpression of the pem gene encoding pectin methylesterase of erwinia chrysanthemi strain 3937.	the pem gene encoding the pectin methylesterase (pme) of erwinia chrysanthemi strain 3937 was subcloned and its nucleotide sequence determined. the gene contains an open reading frame of 1098 bp and codes for a protein of 366 amino acids (aa). the mature 37-kda form of the protein is 342 aa long and has a calculated isoelectric point of 9.64. a plasmid was constructed to overproduce pme: a dna fragment carrying pem was amplified by the polymerase chain reaction and cloned downstream from the pl ...	1993	8370537
[expression of pel genes of erwinia chrysanthemi ena49 in erwinia carotovora var. atroseptica 36a cells].	erwinia atroseptica 36a cells were transformed by the recombinant plasmid ppl5-1 (a derivative of the vector plasmid puc19) containing pelb and pelc genes which encode pectate lyases of erwinia chrysanthemi ena49. synthesis of pectate lyases plb and plc determined by the cloned pel genes is constitutive in erwinia atroseptica 36appl5-1 cells and not inducible by sodium polypectate. the major part of these enzymes was accumulated in the periplasmic fraction of erwinia atroseptica and cells were u ...	1993	8371722
cloning and sequence analysis of a gene (pchr) encoding an arac family activator of pyochelin and ferripyochelin receptor synthesis in pseudomonas aeruginosa.	pseudomonas aeruginosa k372 is deficient in the production of both the 75-kda ferripyochelin receptor protein and pyochelin. a 1.8-kb ecori-sali fragment which restored production of both the receptor protein and pyochelin was cloned. nucleotide sequencing of the fragment revealed an open reading frame of 888 bp, designated pchr (pyochelin), capable of encoding a 296-amino-acid protein of a 32,339-da molecular mass. by using a phage t7-based expression system, a protein of ca. 32 kda was produce ...	1993	8397186
involvement of lipopolysaccharide in the secretion of escherichia coli alpha-haemolysin and erwinia chrysanthemi proteases.	the presence of the alpha-haemolysin secretion genes sensitizes escherichia coli to vancomycin, a glycopeptide antibiotic that is normally excluded from the gram-negative envelope (owing to its large size) (m(r) 1400). the selection of vancomycin mutants in strains carrying such genes was found to be a very powerful method for selecting non-haemolytic mutants. in this way, mutations in the known secretion genes, hlyb, hlyd and tolc, were obtained. however additional mutations mapped in genes rfa ...	1993	8437516
regulation of the expression of a pela::uida fusion in erwinia chrysanthemi and demonstration of the synergistic action of plant extract with polygalacturonate on pectate lyase synthesis.	the phytopathogenicity of erwinia chrysanthemi is chiefly supported by the production of pectate lyase isoenzymes, encoded by the pel genes. one of these enzymes, pela, encoded by the pela gene, seems to represent only a small part of the total pectate lyase activity, but is required for full bacterial pathogenicity. to study the regulation of pela gene expression, a pela::uida gene fusion was constructed. expression of this fusion was analysed under various growth conditions and in various gene ...	1993	8450303
mutagenesis of cellulase egz for studying the general protein secretory pathway in erwinia chrysanthemi.	extracellular secretion of endoglucanase z (egz) from erwinia chrysanthemi is mediated by the so-called out general secretion pathway and, presumably, involves recognition of egz-carried structural information by one or more of the out proteins. investigating the relationships between structure and secretability of egz was the purpose of the present work. egz is made of two independent domains, located at the n- and c-proximal sides, separated by a ser/thr-rich region, which are responsible for ...	1993	8469118
new domain motif: the structure of pectate lyase c, a secreted plant virulence factor.	pectate lyases are secreted by pathogens and initiate soft-rot diseases in plants by cleaving polygalacturonate, a major component of the plant cell wall. the three-dimensional structure of pectate lyase c from erwinia chrysanthemi has been solved and refined to a resolution of 2.2 angstroms. the enzyme folds into a unique motif of parallel beta strands coiled into a large helix. within the core, the amino acids form linear stacks and include a novel asparagine ladder. the sequence similarities ...	1993	8502994
co-expression of an erwinia chrysanthemi pectate lyase-encoding gene (pele) and an e. carotovora polygalacturonase-encoding gene (peh1) in saccharomyces cerevisiae.	a pectate lyase (pl)-encoding gene (pele) from erwinia chrysanthemi and a polygalacturonase (pg)-encoding gene (peh1) from e. carotovora were each inserted between a novel yeast expression-secretion cassette and a yeast gene terminator, and cloned separately into a yeast-centromeric shuttle vector (ycp50), generating recombinant plasmids pams12 and pams13. transcription initiation signals present in the expression-secretion cassette were derived from the yeast alcohol dehydrogenase gene promoter ...	1993	7763727
erwinia chrysanthemi ec16 produces a second set of plant-inducible pectate lyase isozymes.	the enterobacterium erwinia chrysanthemi causes soft-rot diseases involving extensive tissue maceration in a wide variety of plants and secretes multiple pectic enzymes that degrade plant cell walls and middle lamellae. an e. chrysanthemi mutant with directed deletions or insertions in genes pehx, pelx, pela, pelb, pelc, and pele, which encode exo-poly-alpha-d-galacturonosidase, exopolygalacturonate lyase, and four isozymes of pectate lyase, respectively, was constructed by the marker exchange o ...	1993	16348952
two genomic regions involved in catechol siderophore production by erwinia carotovora.	two regions involved in catechol biosynthesis (cbs) of erwinia carotovora w3c105 were cloned by functional complementation of escherichia coli mutants that were deficient in the biosynthesis of the catechol siderophore enterobactin (ent). a 4.3-kb region of genomic dna of e. carotovora complemented the entb402 mutation of e. coli. a second genomic region of 12.8 kb complemented entd, entc147, ente405, and enta403 mutations of e. coli. although functions encoded by catechol biosynthesis genes (cb ...	1994	16349193
ribotyping of erwinia chrysanthemi strains in relation to their pathogenic and geographic distribution.	16s and 23s rrnas from escherichia coli were used to study the relationship among a representative collection of strains of erwinia chrysanthemi differing in their original host and geographical origin. phenetic analysis of restriction fragment length polymorphisms allowed the distribution of the studied strains into seven clusters. these clusters were similar to those obtained by cladistic methods and appeared to correlate well with the established pathovars and biovars but to a lesser extent w ...	1994	16349416
[acute pancreatitis in children with acute lymphoblastic leukemia treated with l-asparaginase].	the treatment of the acute lymphoblastic leukemia in childhood includes frequent administration of l-asparaginase by intravenous route. l-asparaginase is an enzyme produced by e. coli and erwinia chrysanthemi strains. adverse reactions produced by l-asparaginase are numerous, and pancreatitis is being the most severe. children with the acute lymphoblastic leukemia were followed up for 2 years. hyperglycaemia and glycosuria were noted in 10% of them resulting in l-asparaginase cessation or replac ...	1994	7808958
extracellular polysaccharide of erwinia chrysanthemi ech6.	many strains of erwinia chrysanthemi, which are gram-negative bacterial phytopathogens, produce copious amounts of extracellular polysaccharides. the extracellular polysaccharide from e. chrysanthemi pv. zeae strain sr 260, a phytopathogen of corn, is a branched-chain glucomannorhamnan of proven structure (gray et al., carbohydr. res. 1993, 245, 271-287). the extracellular polysaccharide from e. chrysanthemi ech6 is different, containing no rhamnose or mannose. it is composed of l-fucose, d-gala ...	1994	7727344
identification of the integration host factor genes of erwinia chrysanthemi 3937.	two erwinia chrysanthemi homologues of the hima and himd genes of escherichia coli which encode the integration host factor (ihf) were cloned, sequenced and compared to their homolog in other enterobacteria (embl accession nos x74749 and x74750). both genes were inactivated by the insertion of an antibiotic resistance cassette, allowing for the isolation of ihf- mutants of e chrysanthemi.	1994	7748927
expression of the butyrivibrio fibrisolvens endo-beta-1,4-glucanase gene together with the erwinia pectate lyase and polygalacturonase genes in saccharomyces cerevisiae.	recombinant saccharomyces cerevisiae strains capable of simultaneous secretion of bacterial glucanase and pectinase enzymes have been developed. the butyrivibrio fibrrisolvens endo-beta-1,4-glucanase gene (end1), the erwinia chrysanthemi pectate lyase gene (pele) and e. carotovora polygalacturonase gene (peh1) were each inserted between a yeast expression-secretion cassette and yeast gene terminator, and cloned into yeast-centromeric shuttle vectors. transcription initiation signals present in t ...	1994	7750141
the three-dimensional structure of pectate lyase e, a plant virulence factor from erwinia chrysanthemi.	the three-dimensional structure of pectate lyase e (pele) has been determined by crystallographic techniques at a resolution of 2.2 a. the model includes all 355 amino acids but no solvent, and refines to a crystallographic refinement factor of 20.6%. the polypeptide backbone folds into a large right-handed cylinder, termed a parallel [beta] helix. loops of various sizes and conformations protrude from the central helix and probably confer function. a putative ca2+-binding site as well as two ca ...	1994	12232373
erwinia chrysanthemi and pseudomonas syringae: plant pathogens trafficking in extracellular virulence proteins.		1994	7859513
rhizobium meliloti homologs of escherichia coli mur genes.	the pectate-lyase-encoding gene pelb of erwinia chrysanthemi ec16 was used as a probe for hybridization to rhizobium meliloti rm1021 chromosomal dna under low-stringency conditions. an rm1021 dna fragment that hybridized to this probe was cloned and sequenced. results of rna hybridization indicate that a portion of the cloned fragment is transcribed in r. meliloti. although the rm1021 fragment shares no significant nucleotide sequence identity with ec16 pelb, it includes an orf (open reading fra ...	1994	7926844
erwinia chrysanthemi l-asparaginase: epitope mapping and production of antigenically modified enzymes.	this study shows that the antigenicity of erwinia chrysanthemi l-asparaginase can be reduced by site-directed mutagenesis. ten b-cell epitopes of the enzyme were identified using synthetic hexapeptides and polyclonal antisera from rabbits and mice. the region 282givppdeelp292 near the c-terminus was an immunodominant epitope. binding of two hexapeptides (283ivppde288 and 287deelpg292) to the antibodies was dependent on pro285, and pro286, since their replacement by almost any other amino acid re ...	1994	7945221
erwinia chrysanthemi hrp genes and their involvement in soft rot pathogenesis and elicitation of the hypersensitive response.	unlike the bacterial pathogens that typically cause the hypersensitive response (hr) in plants, erwinia chrysanthemi has a wide host range, rapidly kills and macerates host tissues, and secretes several isozymes of the macerating enzyme pectate lyase (pel). pelabce- and out- (secretion-deficient) mutants were observed to produce a rapid necrosis in tobacco leaves that was indistinguishable from the hr elicited by the narrow-host-range pathogens e. amylovora ea321 and pseudomonas syringae pv. syr ...	1994	7949326
verification of elisa results by immunomagnetic isolation of antigens from extracts and analysis with sds-page and western blotting, demonstrated for erwinia spp. in potatoes.	isolation of antigens on immunomagnetic beads and subsequent analysis with sds-page and western blotting (immunomagnetic isolation-western blotting (imi-wb)) was used to verify positive elisa results for erwinia chrysanthemi and erw. carotovora subsp. atroseptica in potato peel extracts. direct analysis of highly contaminated extracts by western blotting without previous immuno-isolation resulted in background reactions, whereas immunomagnetic isolation resulted in distinct bands of specific ant ...	1994	7961189
isolation and characterization of a second exe operon required for extracellular protein secretion in aeromonas hydrophila.	strain c5.84 is a tn5-751 insertion mutant of aeromonas hydrophila which is unable to secrete extracellular proteins, instead accumulating them in the periplasm (b. jiang and s.p. howard, j. bacteriol. 173:1241-1249, 1991). a 3.5-kb bglii fragment which complements this mutation was isolated from the chromosome of the parent strain. analysis of this fragment revealed an operon-like structure with two complete genes, exea and exeb, a functional promoter 5' to the exea gene, and a 13-bp inverted r ...	1994	7961440
prtd, the integral membrane atp-binding cassette component of the erwinia chrysanthemi metalloprotease secretion system, exhibits a secretion signal-regulated atpase activity.	we have overproduced, partially purified, and characterized prtd, the atp-binding cassette (abc) integral membrane component from the metalloproteases secretion system of the gram-negative phytopathogenic bacterium erwinia chrysanthemi. these metalloproteases are secreted independently of the general export pathway encoded by the sec genes. they are secreted via a c-terminal secretion signal and by a secretion apparatus composed of two inner membrane proteins, prtd and prte, and one outer membra ...	1994	7961727
pecs: a locus controlling pectinase, cellulase and blue pigment production in erwinia chrysanthemi.	erwinia chrysanthemi mutants (designated as pecs) displaying derepressed pectate lyase and cellulase synthesis were isolated. in addition, the pecs mutation is responsible for production of an extracellular insoluble blue pigment whose synthesis is cryptic in the wild-type 3937 strain. transduction analysis indicates that the phenotype is due to a single mutation located near the xyl marker on the strain 3937 chromosome. this mutation was complemented by an r-prime plasmid carrying the xyl and a ...	1994	8022282
minimal effects of e. coli and erwinia asparaginase on the coagulation system in childhood acute lymphoblastic leukemia: a randomized study.	a randomized study was done in twenty newly diagnosed children with acute lymphoblastic leukemia. ten children were treated with escherichia coli l-asparaginase, and ten with erwinia chrysanthemi l-asparaginase. l-asparaginase (asp) treatment started halfway during all-induction treatment with vincristine, prednisone, daunorubicin and intrathecal methotrexate. the mean activated partial thromboplastin time (aptt) level in all children demonstrated a significant fall (p < 0.001) from 28.25 sec at ...	1994	8058004
beta-lactamase topology probe analysis of the outo nmephe peptidase, and six other out protein components of the erwinia carotovora general secretion pathway apparatus.	the out gene cluster of erwinia spp. encodes the proteins of the general secretory pathway (gsp) apparatus that is required for pectinase and cellulase secretion. we have used fusions between erwinia carotovora subsp. carotovora (ecc) out genes and the topology probe blam to assess the ability of out protein regions to export blam across the cytoplasmic membrane in escherichia coli and ecc. for the outo gene product (an nmephe peptidase), seven transmembrane regions have been identified and one ...	1994	8065262
secretion of the serratia marcescens hasa protein by an abc transporter.	we previously identified a serratia marcescens extracellular protein, hasa, able to bind heme and required for iron acquisition from heme and hemoglobin by the bacterium. this novel type of extracellular protein does not have a signal peptide and does not show sequence similarities to other proteins. hasa secretion was reconstituted in escherichia coli, and we show here that like many proteins lacking a signal peptide, hasa has a c-terminal targeting sequence and is secreted by a specific atp bi ...	1994	8071214
bglr protein, which belongs to the bglg family of transcriptional antiterminators, is involved in beta-glucoside utilization in lactococcus lactis.	a fragment of the lactococcus lactis chromosome containing an open reading frame of 265 codons, denoted bglr, has been characterized. the polypeptide encoded by bglr shares 36 to 30% sequence identity with a family of regulatory proteins including arbg from erwinia chrysanthemi, bglg from escherichia coli, and sact and sacy from bacillus subtilis. these regulatory proteins are involved in positive control of the utilization of different sugars by transcription antitermination. for some of these ...	1994	8083160
specific interactions of erwinia chrysanthemi kdgr repressor with different operators of genes involved in pectinolysis.	the erwinia chrysanthemi kdgr gene encodes a repressor that negatively regulates the expression of genes involved in pectinolysis and in pectinase secretion. the cloned kdgr gene was overexpressed in escherichia coli by using a phage t7 system. overproduced repressor was purified to homogeneity by two chromatographic steps. gel retardation and dnase i protection experiments demonstrated the specific binding of the kdgr protein to the operators of pectinase genes (pela, pelb, pelc, pele), to the ...	1994	8107132
role of endoglucanases in erwinia chrysanthemi 3937 virulence on saintpaulia ionantha.	the role of endoglucanases (endoglucanases z and y) in erwinia chrysanthemi pathogenicity on saintpaulia ionantha was assessed by mutagenizing cloned cel genes (celz and cely) and recombining them with the chromosomal alleles. strains with an omega interposon in celz, a deletion in cely, or a double cel mutant were as virulent as the wild-type strain. however, in the strain with a deletion in cely, a delay in the appearance of symptoms was observed, and then maceration progressed as in plants in ...	1994	8113196
effect of mannitol crystallinity on the stabilization of enzymes during freeze-drying.	the stabilizing effect of mannitol during the freeze-drying of proteins was studied using l-lactate dehydrogenase (ldh, rabbit muscle), beta-galactosidase (escherichia coli) and l-asparaginase (erwinia chrysanthemi) as model proteins. crystallization of mannitol was studied by powder x-ray diffraction and differential scanning calorimetry (dsc), in relation to the stabilizing effect. all the enzymes were protected concentration-dependently by amorphous mannitol, but the stabilizing effect was de ...	1994	8124765