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production of cellulase (cx) by different species of erwinia.the tested isolates of erwinia chrysanthemi (corn pathotype) and e. carotovora constitutively produce high levels of cellulase(s) (cx) in presence or absence of carboxymethyl-cellulose (cmc) as substrate in the medium. the tested isolates of e. atroseptica produced high levels of cellulase when grown in presence of both carboxymethyl-cellulose (cmc) and sucrose, low levels in presence of carboxymethyl-cellulose alone, and traces of cellulase(s) in presence of sucrose alone. the activity of cellu ...197938608
taxonomy of the genus serratia.one hundred and fifty-six strains of serratia and related bacteria including representatives of enterobacter liquefaciens, enterobacter cloacae, enterobacter aerogenes, erwinia carotovora, erwinia chrysanthemi, erwinia herbicola and erwinia nimipressuralis were studied using 223 morphological, physiological, biochemical and carbon source utilization tests. the results were subjected to computer analysis. at the 80% similarity level all strains, except two, grouped into eight phenons representing ...1977319202
lysinoalanine utilization by erwinia chrysanthemi and escherichia coli. 1979387557
bacterial stalk rot of maize, its symptoms and host-range.stalk rot of maize, caused by erwinia carotovora f. sp. zeae sabet (re-designated as pectobacterium chrysanthemi pathovar. zeae by kelman 1974) showed first premature withering and drying up of the uppermost leaves which was soon followed by the lower leaves. the rot either extended from the base upwards (basal rot) or from the top downwards (top rot). in the case of basal rot, the leaves become yellow and the infected tissue becomes brown, soft, and water soaked. internally, the stalk turns int ...1977857508
[study of the drug resistance of bacteria of the genus erwinia].sensitivity of 788 strains of erwinia isolated from natural substrates and received from other laboratories was studied with respect to penicillin, streptomycin, tetracycline, chloramphenicol and kanamycin. it was found that 346 (43.9 per cent) strains were drug resistant. however, the levels of the resistance to most of the drugs tested were not high and amounted to 5-10 gamma/ml. penicillin resistance was the most frequent characteristics of the strains studied (341 strains). the resistance le ...1977883805
donor strains of the soft-rot bacterium erwinia chrysanthemi and conjugational transfer of the pectolytic capacity.donor strains of erwinia chrysanthemi icpb ec16, a member of the soft-rot (pectolytic) section of the enterobacterial genus erwinia, were obtained by chromosomal integration of an f'lac(+) plasmid originating from escherichia coli. these stable donor strains, selected from an unstable f'lac(+) heterogenote by repeated platings of single lac(+) colonies on lactose minimal agar, do not segregate (as does the parent f'lac(+) heterogenote) into lac(-) or f(-) clones, in either the presence or absenc ...1977924974
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 ...19921299839
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 ...19921319773
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 ...19921333438
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 ...19911367534
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 ...19921368993
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 ...19921392469
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 ...19921406275
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 ...19921427097
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 ...19921427098
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 ...19921429461
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 ...19921447147
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 ...19921453958
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 ...19921494344
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 ...19921508046
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 ...19921545703
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 ...19921545707
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 ...19921545709
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 ...19921548242
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 ...19921551853
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 = ...19921563515
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 ...19921593262
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 ...19921622220
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 ...19921624465
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 ...19921629152
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 ...19911657869
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 ...19911660698
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 ...19911662754
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 ...19911677466
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 ...19921729230
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 ...19921732212
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 ...19911766386
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 ...19911787788
[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 ...19911787841
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 ...19911791757
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 ...19911829728
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 ...19911840643
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 ...19911868195
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 ...19911874406
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 ...19911904127
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 ( ...19911917878
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 ...19911937031
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 ...19911992458
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 ...19912007544
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 ...19912045776
secretion processing and activation of erwinia chrysanthemi proteases.e chrysanthemi, a phytopathogenic enterobacterium, secretes several enzymes into the medium such as pectinases cellulases and proteases. it also produces 3 distinct and antigenically related extracellular proteases. the proteases secretion pathway seems to be distinct from that of the other extracellular enzymes since pleiotropic mutants impaired in cellulase and pectinase secretion are unimpaired in protease secretion. e chrysanthemi proteases b and c secretion occurs without an n-terminal sign ...19902116182
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 ...19902126210
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 ...19902155205
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 ...19902160934
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 ...19902168372
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 ...19902184029
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 ...19902188956
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.19902195018
[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 ...19902199828
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 ...19902211513
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 ...19902211614
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 ...19902228958
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, ...19902233523
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 ...19902254266
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 ...19902279995
expanded linkage map of erwinia chrysanthemi strain 3937.in this paper we describe the chromosomal location of various loci in erwinia chrysanthemi strain 3937. auxotrophic markers were obtained by chemical mutagenesis, antibiotic resistances were isolated spontaneously and mutations in sugar utilization were obtained by means of mu insertions. these markers were located on the genetic linkage map of strain 3937 by using a conjugative system mediated by rp4::mini-mu plasmids which permitted transfer of genetic material from any point of origin. the lo ...19892527330
cloning and analysis of structural and regulatory pectate lyase genes of erwinia chrysanthemi ena49.erwinia chrysanthemi ena49 structural and regulatory ptl genes, coding for pectate lyase (ptl) were cloned in escherichia coli cells. phage vector lambda l47.1 and phasmid vector lambda pmyf131 were used for constructing libraries of bamhi and ecori fragments, respectively, of er. chrysanthemi chromosomal dna. among the 1,100 hybrid clones containing bamhi er. chrysanthemi dna fragments and 11,000 hybrid clones containing ecori fragments, six and 45 clones, respectively, were identified as havin ...19892530137
nucleotide sequence of an erwinia chrysanthemi gene encoding shikimate kinase. 19892537963
molecular cloning of the outj gene involved in pectate lyase secretion by erwinia chrysanthemi.the phytopathogenic enterobacterium erwinia chrysanthemi secretes a number of enzymes involved in plant-tissue degradation, notably the five isoenzymes of pectate lyase. we have cloned a region involved in pectate lyase and cellulase secretion by complementation of non-secretory outj mutants of e. chrysanthemi strain 3937 using the rp4::minimu plasmid pulb110. the cloned region maps near the ade-22 marker on the e. chrysanthemi 3937 chromosome. an r-prime containing a chromosomal dna insert of a ...19892546003
sequence analysis of the clostridium cellulolyticum endoglucanase-a-encoding gene, celcca.the nucleotide sequence of a clostridium cellulolyticum endo-beta-1,4- glucanase (egcca)-encoding gene (celcca) and its flanking regions, was determined. an open reading frame (orf) of 1425 bp was found, encoding a protein of 475 amino acids (aa). this orf began with an atg start codon and ended with a taa ochre stop codon. the n-terminal region of the egcca protein resembled a typical signal sequence of a gram-positive bacterial extracellular protein. a putative signal peptidase cleavage site w ...19892558058
relationship between the pel genes of the pelade cluster in erwinia chrysanthemi strain b374.in this paper, we have used filter hybridization and nucleotide sequencing to analyse the relationship between the three genes of the pelade cluster in the erwinia chrysanthemi (ech) strain b374. this cluster encodes for three of the five pectate lyase proteins that are involved in the maceration and soft-rotting of plant tissue, an important trait in ech pathogenicity. southern hybridization revealed homology between each of the three pel genes. a 3560 bp dna fragment containing the pele and pe ...19892615652
characterization of a protein inhibitor of extracellular proteases produced by erwinia chrysanthemi.erwinia chrysanthemi, a phytopathogenic bacterium, produces a protease inhibitor which is a low-molecular-weight, heat-stable protein. in addition to its action on the three e. chrysanthemi extracellular proteases a, b and c, it also strongly inhibits the 50 kd extracellular protease of serratia marcescens. its structural gene (inh) was subcloned and expressed in escherichia coli, in which it encodes an active inhibitor which was purified. the nucleotide sequence of the inh gene shows an open re ...19892654540
[reca-genes of erwinia chrysanthemi ena49].the reca gene of erwinia chrysanthemi ena49 has been cloned in vivo in escherichia coli k12, reca13 cells using the plasmid pulb113. on the basis or dna repair and recombination deficiencies complementation, of restoration of the inducible "sos"-response functions the functional identity of the cloned gene with the reca gene was concluded. the reca gene was localized in the 18th min region of the chromosomal genetical map of erwinia chrysanthemi ena49 between the genes proa and phea.19892659980
nucleotide sequence of the erwinia chrysanthemi gene encoding 2-keto-3-deoxygluconate permease.the phytopathogenic bacterium erwinia chrysanthemi produces a group of pectolytic enzymes able to depolymerise the pectic compounds in plant cell walls. the resulting tissue maceration is known as soft rot disease. the degraded pectin products are transported by 2-keto-3-deoxygluconate permease into the bacterial cell, where they serve as carbon and energy sources. this h+ coupled transport system is encoded by the kdgt gene; we report the nucleotide sequence of kdgt. it is encoded by an open re ...19892684787
isolation of erwinia chrysanthemi mutants altered in pectinolytic enzyme production.various mutations in the pectin catabolic pathway of erwinia chrysanthemi were isolated by selection of mu-lac insertions, resulting in expression of the lac genes inducible by pectin degradation products. this approach allowed us to isolate lacz fusions with the genes pelc, peld, ogl and pem, encoding pectate lyases plc and pld, oligogalacturonate lyase and pectin methylesterase, respectively. moreover, we obtained mutations affecting the regulation of pectinolytic enzymes; a locus called pecl ...19892693903
nucleotide sequences of the erwinia chrysanthemi ogl and pele genes negatively regulated by the kdgr gene product.the nucleotide sequences of the coding and regulatory regions of the genes encoding oligoglacturonate lyase (ogl) and pectate lyase e isoenzyme (ple) from erwinia chrysanthemi 3937 were determined. the ogl sequence contains an open reading frame (orf) of 1164 bp coding for a 388-amino acid (aa) polypeptide with a predicted mr of 44,124. a possible transcriptional start signal showing homology with the escherichia coli promoter consensus sequence was detected. in addition, a sequence 3' to the co ...19892695393
the beta-glucosides metabolism in erwinia chrysanthemi: preliminary analysis and comparison to escherichia coli systems. 19892699245
escherichia coli molybdoenzymes can be activated by protein fa from several gram-negative bacteria.six gram-negative bacteria (klebsiella pneumoniae, erwinia chrysanthemi, proteus vulgaris, serratia marescens, salmonella typhimurium, and pseudomonas aeruginosa) were shown to contain an fa-type protein capable of activating aponitrate reductase, apotrimethylamine n-oxide reductase and apoformate dehydrogenase of escherichia coli. protein fa activity was highest in erwinia chrysanthemi and lowest in pseudomonas aeruginosa. all the species also contained the low-mr (less than or equal to 1500) h ...19892699894
protease secretion by erwinia chrysanthemi. proteases b and c are synthesized and secreted as zymogens without a signal peptide.the gene encoding the secreted 53-kda metalloprotease (protease b) and the 5' end of the gene encoding the secreted 55-kda metalloprotease (protease c) of the gram-negative bacterium erwinia chrysanthemi have been sequenced. the predicted sequences of the two proteases do not have typical signal sequences at their nh2 termini. both proteases are synthesized as inactive higher molecular weight precursors (zymogens prob and proc) which are secreted into the external medium where divalent cation-me ...19892722818
preliminary crystallographic analysis of a plant pathogenic factor: pectate lyase.pectate lyase is a saccharide-binding enzyme that lyitically depolymerizes polypectate in higher plant cell walls, thus causing soft-rot diseases in food crops. a pectate lyase from erwinia chrysanthemi, ec16 (ple), crystallizes in the orthorhombic space group p2(1)2(1)2(1) with unit cell dimension of a = 39.0 a, b = 91.0 a and c = 103.4 a. the asymmetric unit consists of one molecule with a molecular mass of 38,118 daltons and the x-ray diffraction extends to a resolution of 1.8 a. the crystals ...19892769765
cellulase families revealed by hydrophobic cluster analysis.the amino acid sequences of 21 beta-glycanases have been compared by hydrophobic cluster analysis. six families of cellulases have been identified on the basis of primary structure homology: (a) endoglucanases b, c and e of clostridium thermocellum; endoglucanases of erwinia chrysanthemi and bacillus sp.; endoglucanase iii of trichoderma reesei; endoglucanase i of schizophyllum commune; (b) cellobiohydrolase ii of t. reesei; endoglucanases of cellulomonas fimi and streptomyces sp; (c) cellobiohy ...19892806912
[secretion of pectate lyase and protein composition of the outer membrane of erwinia chrysanthemi ena49 cells].the combined changes (specters of isoenzymes of intracellular and extracellular pectatelyase, protein composition of periplasm and outer membrane) in the cells of e. chrysanthemi ena49 from the periodical culture growing on the inducer containing medium have been studied. the beginning of active pectatelyase synthesis was established to be accompanied by the temporal intracellular accumulation of the enzyme. the cellular capability of pectatelyase secretion to the outer medium correlates with th ...19892811908
characterization of erwinia chrysanthemi extracellular proteases: cloning and expression of the protease genes in escherichia coli.erwinia chrysanthemi, a phytopathogenic enterobacterium, secretes three antigenically and structurally distinct proteases, a, b, and c and produces a protease inhibitor, a low-molecular-weight, heat-stable protein which remains mostly intracellular and which binds specifically to the a, b, and c proteases. the structural genes for proteases a, b, and c and for the inhibitor are clustered on a ca. 40-kilobase dna fragment present in cosmid pew4. escherichia coli strains harboring pew4 secrete the ...19872822661
molecular cloning and sequencing of a pectate lyase gene from yersinia pseudotuberculosis.a pectate lyase gene (pely) from yersinia pseudotuberculosis was cloned in escherichia coli dh-5 alpha. the gene was expressed in either orientation in puc plasmids, indicating that the insert dna carried a y. pseudotuberculosis promoter which functioned in e. coli. however, when cloned in the orientation which placed the coding region downstream of the vector lac promoter, expression of pely was nine times higher than it was in the opposite orientation and the growth of e. coli cells was inhibi ...19882832382
homology between endoglucanase z of erwinia chrysanthemi and endoglucanases of bacillus subtilis and alkalophilic bacillus.nucleotide sequencing of the celz gene encoding the extracellular endoglucanase z of erwinia chrysanthemi indicated the presence of an open reading frame encoding 428 amino acids. the mature protein appeared to be extended by a signal peptide of 43 amino acids; this sequence is unusually long and positively charged (+5). it was shown to function as a signal peptide by fusing it to a truncated phoa gene encoding escherichia coli alkaline phosphatase. comparison of the encoded sequence with those ...19882835589
molecular cloning and nucleotide sequence of the pectin methyl esterase gene of erwinia chrysanthemi b374.the gene encoding pectin methyl esterase (pme) has been cloned from erwinia chrysanthemi b374. expression of pme in escherichia coli allowed the enzyme to be characterized. pectin methyl esterase (pme) was found to have an apparent molecular weight of 36,000 daltons and an isoelectric point of approximately 9.9. the structural gene was sequenced and consists of a 1098-bp open reading frame encoding a polypeptide of 39,318 daltons, which includes an amino-terminal signal peptide. the isolation of ...19882837615
chromosomal mapping of the pel and cel genes in erwinia chrysanthemi strain b374.using the rp4::mini-mu in vivo cloning technique, van gijsegem et al. (1985) isolated several pel and cel genes of erwinia chrysanthemi (ech) b374 strain. we have localized these genes on the ech chromosome by co-transfer mapping of mudi1734 insertion mutants and refined the map by co-transposition analysis. this analysis has enabled us to identify another cel gene.19882837618
isolation, characterization, and synthesis of chrysobactin, a compound with siderophore activity from erwinia chrysanthemi.a catechcol-type siderophore, assigned the trivial name chrysobactin, was isolated from the phytopathogenic bacterium erwinia chrysanthemi and characterized by degradation and spectroscopic techniques as n-[n2-(2,3-dihydroxybenzoyl)-d-lysyl]-l-serine. chrysobactin, which was also obtained by chemical synthesis, was shown to be active in supplying iron to a group of mutants of e. chrysanthemi defective in biosynthesis of the siderophore.19892914949
amber suppressors of erwinia chrysanthemi.mutations trp1 and thya1, both of a polyauxotrophic derivative of the erwinia chrysanthemi strain b374, were characterized as amber mutations with an escherichia coli suppressor, supa1p2, which inserts a glutamine in response to uag. simultaneous reversion of both mutations allowed us to isolate amber suppressor mutants of e. chrysanthemi. these suppressors were tested with a set of amber mutants of bacteriophage mu which had been previously characterized on e. coli. the two independently isolat ...19872956976
mapping and regulation of the cel genes in erwinia chrysanthemi.chromosomal mutations of the celz and cely genes which encode two different endoglucanases in erwinia chrysanthemi 3937 were obtained by a three-step procedure: (i) in escherichia coli, insertions of lacz fusion-forming mini-mu bacteriophages in the cel genes cloned on plasmids and screening of cel-lac fusions, (ii) mu-mediated transduction in e. chrysanthemi of the plasmids carrying the fusions, (iii) recombinational exchange between the plasmidic mutated and the wild-type chromosomal alleles. ...19882963946
molecular cloning in escherichia coli of erwinia chrysanthemi genes encoding multiple forms of pectate lyase.the phytopathogenic enterobacterium erwinia chrysanthemi excretes multiple isozymes of the plant tissue-disintegrating enzyme, pectate lyase (pl). genes encoding pl were cloned from e. chrysanthemi cucpb 1237 into escherichia coli hb101 by inserting sau3a-generated dna fragments into the bamhi site of pbr322 and then screening recombinant transformants for the ability to sink into pectate semisolid agar. restriction mapping of the cloned dna in eight pectolytic transformants revealed overlapping ...19852982794
pulb113, an rp4::mini-mu plasmid, mediates chromosomal mobilization and r-prime formation in erwinia amylovora, erwinia chrysanthemi, and subspecies of erwinia carotovora.the rp4::mini-mu plasmid pulb113, transferred from escherichia coli strain mxr, was stable and transfer proficient in erwinia amylovora strain ea303, e. carotovora subsp. atroseptica strain eca12, e. carotovora subsp. carotovora strain ecc193, and e. chrysanthemi strain ec183. the plasmid mobilized an array of erwinia sp. chromosomal markers (e. amylovora: his+,ilv+,rbs+,ser+,thr+;e. chrysanthemi:arg+,his+,ilv+,leu+; e. carotovora subsp. atroseptica: arg+,gua+,leu+,lys+,pur+,trp+; e. carotovora ...19852992373
mu-lac insertion-directed mutagenesis in a pectate lyase gene of erwinia chrysanthemi.the pelc gene, which encodes one of the five major pectate lyase (pl) isoenzymes in erwinia chrysanthemi 3937, designated plc, was subcloned from a hybrid lambda phage into a pbr322 derivative and mutagenized with a mini-mu-lacz transposable element able to form fusions to the lacz gene. one plasmid (pad1) which had an inactivated pelc gene and a lac+ phenotype was selected in escherichia coli. this plasmid was introduced into erwinia chrysanthemi, and the pelc::mini-mu insertion was substituted ...19852993251
marker-exchange mutagenesis of a pectate lyase isozyme gene in erwinia chrysanthemi.the phytopathogenic enterobacterium erwinia chrysanthemi contains pel genes encoding several different isozymes of the plant-tissue-disintegrating enzyme pectate lyase (pl). the pelc gene, encoding an isozyme with an approximate isoelectric point of 8.0, was mutagenized by a three-step procedure involving (i) insertional inactivation of the cloned gene by ligation of a kan-containing bamhi fragment from puc4k with a partial sau3a digest of e. chrysanthemi pelc dna in pbr322; (ii) mobilization of ...19852995324
[the role of te transposition of tn1000 from flac+-plasmid into chromosome of erwinia chrysanthemi in the formation of hfr-type donors].based on the data of stability of the donor state of hfr-like strain erwinia chrysanthemi vy1-10 in reca+ and reca- cells, it can be suggested that the donor properties of the strain are mediated by the presence of the genetic homology region which occurred as a result of transposition of the tn1000 from the flac+ plasmid into the chromosome of e. chrysanthemi. tn1000 may be transposed into several sites on the chromosome of e. chrysanthemi ena49. this leads to the appearance of donors transferr ...19853000870
single-site chromosomal tn5 insertions affect the export of pectolytic and cellulolytic enzymes in erwinia chrysanthemi ec16.exponentially growing cells of erwinia chrysanthemi ec16 usually export about 98% of their pectate lyase (pl) and protease, about 40% of their polygalacturonase (pg), and about 60% of their cellulase (endoglucanase or carboxymethyl cellulase; cl). by using the r plasmid, pjb4ji (pph1ji::mu::tn5), three independent tn5 insertion mutants were obtained that exported normal levels of protease but 10% or less of pl, pg, and cl. physical analysis revealed that single copies of tn5 had inserted into th ...19853002271
influence of gyra mutation on expression of erwinia chrysanthemi clb genes cloned in escherichia coli.erwinia chrysanthemi clb genes cloned into nals escherichia coli allowed growth on cellobiose, arbutin, or salicin. in contrast, nalr isogenic strains grew only on cellobiose. it is proposed that expression of cloned e. chrysanthemi clb genes is reduced by the e. coli chromosomal gyra (nalr) mutation, resulting in apparent segregation of the clb and arb sal characters.19863007437
cloning and expression of the erwinia chrysanthemi asparaginase gene in escherichia coli and erwinia carotovora.a genomic library of erwinia chrysanthemi dna was constructed in bacteriophage lambda 1059 and recombinants expressing er. chrysanthemi asparaginase detected using purified anti-asparaginase igg. the gene was subcloned on a 4.7 kb ecori dna restriction fragment into puc9 to generate the recombinant plasmid pasn30. the position and orientation of the asparaginase structural gene was determined by subcloning. the enzyme was produced at high levels in escherichia coli (5% of soluble protein) and wa ...19863011958
evidence of homology between the pectate lyase-encoding pelb and pelc genes in erwinia chrysanthemi.the genes for two of several pectate lyase isozymes produced by the phytopathogenic enterobacterium erwinia chrysanthemi 1237 were subcloned and compared by dna-dna hybridization, and the encoded proteins were analyzed. the borders of the genes were located on a restriction map by incremental exonuclease iii deletions. dna-dna hybridization studies revealed a low percentage of mismatch (7 to 17%) between pelb and pelc. no homology was detected between pelc and other regions of the e. chrysanthem ...19863013832
lactose and melibiose metabolism in erwinia chrysanthemi.a lac+ mutant of erwinia chrysanthemi was isolated from the lac- wild type on lactose agar. beta-galactosidase was expressed independently of lactose transport in both the mutant and the wild type, and neither strain expressed thiogalactoside transacetylase. lactose transport and alpha-galactosidase, constitutive in the lac+ strain, were coordinately induced in the lac- strain by melibiose and raffinose but not by isopropyl-beta-d-thiogalactopyranoside or thiomethyl-beta-d-galactopyranoside. mel ...19863023289
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