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[biological and physicochemical properties of virulent and temperate phages of erwinia carotovora]. 19807382866
[certain features of ornithine decarboxylase biosynthesis by erwinia carotovora 438]. 19807402100
[study of the transfecting properties of the dna of virulent and moderate erwinia carotovora phages]. 19807402109
studies on the preservation of erwinia carotovora var. chrysanthemi (burkholder et al.) dye cultures.it can be concluded that the maize stalk rot pathogen erwinia carotovora var. chrysanthemi (burkholder et al.). dye retained its viability and infectivity for three years when kept in sterile distilled water at low temperature.19807424221
[physicochemical properties of the dna of virulent and moderate erwinia carotovora phages]. 19807432206
sensitivity of some erwinia carotovora serogroups to macromolecular bacteriocins.representative strains from each of 18 erwinia carotovora serogroups were tested for bacteriocin activity. eight bacteriocin producing strains were found and the bacteriocins partially purified by ammonium sulfate fractionation and high-speed centrifugation. bacteriocins from all eight strains were morphologically similar to bacteriophage tails. specific absorption of bacteriocins from one of the antagonistic strains to sensitive bacterial cells was demonstrated with electron microscopy. in four ...19807459719
[localization of ornithine decarboxylase in the cell of erwinia carotovora 438]. 19807464573
phosphatidylethanolamine distribution and fluidity in outer and inner membranes of the gram-negative bacterium erwinia carotovora.1. the distribution of phosphatidylethanolamine, the major lipid of erwinia carotovora, was investigated in intact bacteria, spheroplasts and outer- and inner-membrane preparations, with the amino-group reagent 2,4,6-trinitrobenzenesulphonic acid. only 4% was found on the external surface of the outer membrane with 30% on the internal surface, whereas the inner membrane had 27 and 38% on its external and internal surfaces respectively. some comparative studies were made with three other bacteri ...19806996674
improvement in the persistence of microbial asparaginase and glutaminase in the circulation of the rat by chemical modifications.three enzymes used in cancer chemotherapy (asparaginases from escherichia coli and erwinia carotovora and glutaminase from achromobacter) were each reacted with four amino specific reagents (ethyl acetimidate, o-methylisourea, succinic anhydride, and formaldehyde/sodium borohydride). the half-lives of the modified enzymes measured in the blood of rats showed that guanidation, acetimidation and reductive alkylation were more likely to increase the persistence of the native enzymes than succinylat ...19816794649
plasmid colvbtrp maintenance in erwinia carotovora.plasmid colvbtrp maintenance in erwinia carotovora cells was followed by measuring kinetics of elimination of plasmid genetic markers and loss of plasmid deoxyribonucleic acid. an e. carotovora mutant stably carrying plasmid colvbtrp was isolated. besides stable plasmid maintenance, the mutant showed altered sensitivity to male-specific phage ms2, sensitivity to drugs, and colony morphology.19816944305
[enterobacterial taxonomy: erwinia carotovora and yersinia enterocolitica].the immunotyping of the intracellular protein complex in enterobacteria has allowed to reveal that the intracellular proteins of e. carotovora and y. enterocolitica possess an antigenic profile characteristics of bacteria belonging to the family enterobacteriaceae. e. carotovora are similar to the main group of enterobacteria (escherichia, shigella, salmonella, enterobacter, klebsiella, citrobacter, arizona) in the degree of the antigenic homology of their intracellular protein complex. y. enter ...19817293572
host-pathogen interactions : xviii. isolation and biological activity of glycinol, a pterocarpan phytoalexin synthesized by soybeans.a previously unrecognized phytoalexin has been isolated from soybean cotyledons that had been infected with bacteria or exposed to ultraviolet light. the phytoalexin has been purified to homogeneity by silica gel flash chromatography and high pressure liquid chromatography. it has been structurally characterized by its ultraviolet, circular dichroism and nuclear magnetic resonance spectra, polarimetry, and its mass spectrometric fragmentation pattern. the phytoalexin, (6as,11as)-3,6a,9-trihydrox ...198116661917
[physicochemical and biological properties of the dna of virulent and temperate erwinia carotovora phages].dna of virulent and moderate e. carotovora viruses, of marked at-type, was shown to have molecular weights determine by 2 independent methods from 49.2 to 54 and 28 to 29.8 md, respectively. the genomes of the viruses under study differed by the point and curve profiles of the melting temperature and sedimentation constant. dna of the moderate phage contains an unidentified abnormal base and has no free cohesive ends. dna of the moderate virus contains 15% of nucleotide sequences homologous with ...19827072231
[properties of a toxic substance from erwinia carotovora var. citrullis].the toxic substance of erwinia carotovora, isolated in a pure form, is a protein-polysaccharide complex with molecular weight of 90000 daltons. the protein part of the complex contains 17 amino acids with the prevalence of dicarbonic acids and with insignificant quantities of thyrosine and phenylalanine, while the polysaccharide part of the complex contains 6 sugars. the antigenic specificity of the complex is linked with its hydrocarbon part, and the toxicity of the complex is determined by pro ...19827080758
immunological and pharmacological characterization of poly-dl-alanyl-modified erwinia carotovora l-asparaginase.the covalent attachment of poly-dl-alanine peptides to lysyl residues on the surface of erwinia carotovora l-asparaginase has produced a modified enzyme which is much less immunogenic in mice and demonstrates 100-fold longer plasma half-life in the rhesus monkey. immunogenic responses towards both the immunoglobulin g (igg) and immunoglobulin e (ige) antibody subclasses were evaluated in c57bl x dba/2 f1 mice exposed to 250 rads of whole-body irradiation 4 hr prior to immunization with 5-diazo-4 ...19827105004
[electron microscopic structure of erwinia carotovora moderate phage 59 and its dna].electron microscopic examinations showed the moderate phage 59 erwinia carotovora to consist of a head of icosahedral shape 50.0+/- +/-2.5 nm in diameter, a noncontractile process 150.0+/-3.5 nm in length and 8.2+/-1.0 nm in diameter with a basal plate of 20.0+/-1,5 nm. the head of the virion of 45.9 x 10(3) nm3 in volume is formed by 812 morphological subunits about 3.0 nm in diameter; the number of structural subunits is 4860 on the basis of triangulation number (81). the process of the virus ...19827123916
[isolation and characteristics of a new temperate phage ec-3 of erwinia carotovora]. 19827154993
utilization of a thermosensitive episome bearing transposon tn10 to isolate hfr donor strains of erwinia carotovora subsp. chrysanthemi.a thermosensitive episome bearing the transposon tn10, f(ts)::tn10 lac+, has been successfully transferred from escherichia coli to several wild strains of the enterobacteria erwinia carotovora subsp. chrysanthemi, which are pathogenic on saintpaulia ionantha. in one of these strains, all of the characters controlled by this episome (lac+, tetr, tra+) were expressed, and its replication was stopped at 40 degrees c and above. at 30 degrees c, the episome was easily transferred between strains der ...19826277860
direct dna repeat in plasmid r68.45 is associated with deletion formation and concomitant loss of chromosome mobilization ability.plasmid r68.45 has been useful in promoting the transfer of chromosomal markers in bacteria of many genera. in donors harboring r68.45, chromosome-mobilizing ability (cma) may be lost without the loss of other plasmid markers. however, cma can be somewhat stabilized by maintenance of the donors in the presence of kanamycin (km). we isolated variants of r68.45 from four bacterial species of three genera. plasmid variants isolated included those without cma, without transfer function (tra) and cma ...19826277862
the chromosomal origin of replication (oric) of erwinia carotovora.the chromosomal dna replication origin (oric) of the plant pathogen erwinia carotovora has been isolated and sequenced. the minimal e. carotovora oric regional functional in escherichia coli is a 374 base pair region located on a 7.9 kilobase pair sali fragment which also contains a functional asna gene. differences between the nucleotide sequence of the minimal origin regions of e. carotovora and those of e. coli and salmonella typhimurium are clustered nucleotide substitutions, with regions of ...19826281738
[localization of putrescine in erwinia carotovora atcc 15713 cells]. 19836336577
[isolation of mutants of the erwinia carotovora temperate phage 59 under the action of n-methyl-n'-nitro-n-nitrosoguanidine]. 19836400791
chromosome transfer and r-prime plasmid formation mediated by plasmid pulb113 (rp4::mini-mu) in alcaligenes eutrophus ch34 and pseudomonas fluorescens 6.2.plasmid pulb113 (rp4::mini-mu), which contains the mini-mu transposon, promoted both homologous and heterologous gene transfer from pseudomonas fluorescens 6.2 and alcaligenes eutrophus ch34. homologous gene transfer in p. fluorescens 6.2 and a. eutrophus ch34 occurred at a frequency of 10(-4) to 10(-5), and recombinants inherited unselected recessive markers, suggesting a process of chromosome mobilization. loci involved in autotrophic growth were among those transferred in a. eutrophus. in het ...19836411681
utilization of plasmid pulb113 (rp4::mini-mu) to construct a linkage map of erwinia carotovora subsp. chrysanthemi.we report experimental evidence that pulb113, an rp4::mini-mu plasmid, mediates chromosome transfer in a strain of erwinia carotovora subsp. chrysanthemi which does not accept the f episome. this allowed us to construct a genetic map of that strain by measuring the frequencies of cotransfer of different markers (thy, leu, pro, [his, trp], thya, rpsl, ile).19836574127
the effect of freeze-drying on the quaternary structure of l-asparaginase from erwinia carotovora.l-asparaginase (l-asparagine amidohydrolase, ec 3.5.1.1) from erwinia carotovora undergoes extensive dissociation from active tetramer to inactive monomers when freeze-dried. the monomeric state is stabilized by reconstitution of the freeze-dried enzyme with buffers of high ph and high ionic strength. some compounds, particularly sugars and sugar derivatives, prevent dissociation on freeze-drying, whereas others, such as urea and chaotropic ions, increase dissociation. the effects of additives a ...19836652094
fluoride-induced filaments of erwinia carotovora.sodium fluoride induces filamentous growth of erwinia carotovora when it is grown in liquid media containing aspartic acid only as the sole source of nitrogen. it is proposed that a stable complex of f(-)-mg2+-enzyme and po2(4) resulting in mg2+ deficiency and consequent inability of e. carotovora cells to oxidize aspartic acid normally, is responsible for the formation of filaments in the presence of fluoride ions.19836684838
aldohexuronate transport system in erwinia carotovora.the biochemical and physiological aspects of hexuronate transport in erwinia carotovora were studied to approach the genetic regulation of the hexuronate degradative pathway in this bacterial species. an active transport system for glucuronate and galacturonate uptake exists in e. carotovora. the glucuronate entry reaction displayed saturation kinetics with an apparent km of 0.05 mm (at 25 degrees c; ph 7). galacturonate appeared to be a competitive inhibitor of glucuronate uptake with a ki of 0 ...19836841313
in vivo cloning of erwinia carotovora genes involved in the catabolism of hexuronates.using the rp4::mini-mu pulb113 plasmid, an rp4 derivative carrying a deleted mu prophage which allows the plasmid to pick up any chromosomal dna segment to form r' plasmids, we cloned all of the genes of erwinia carotovora involved in the catabolism of the hexuronates and in the transport of these substrates. with the r' plasmids we isolated, we performed complementation analysis and found that, in the erwinia carotovora strain we used, the genes involved in the catabolism of the hexuronates are ...19836853444
structure of erwinia carotovora temperate bacteriophage 59 and its dna.the temperate phage 59 from erwinia carotovora and its dna were studied. the phage particles have an icosahedral head and a long noncontractile tail with a base plate. the virus dna makes up 50% of the total virus and exists as a linear molecule (molecular weight, 2.6 x 10(7)). a model of virus structural organization is presented.19836854737
host-pathogen interactions : xxiii. the mechanism of the antibacterial action of glycinol, a pterocarpan phytoalexin synthesized by soybeans.the biochemical basis for the ability of the pterocarpan phytoalexin glycinol (3,6a,9-trihydroxypterocarpan) to inhibit the growth of bacteria was examined. glycinol at bacteriostatic concentrations (e.g. 50 micrograms per milliliter) inhibits the ability of erwinia carotovora to incorporate [(3)h]leucine, [(3)h]thymidine, or [(3)h]uridine into biopolymers. exposure of escherichia coli membrane vesicles to glycinol at 20 micrograms per milliliter results in inhibition of respiration-linked trans ...198316663042
host-pathogen interactions : xxv. endopolygalacturonic acid lyase from erwinia carotovora elicits phytoalexin accumulation by releasing plant cell wall fragments.heat-labile elicitors of phytoalexin accumulation in soybeans (glycine max l. merr. cv wayne) were detected in culture filtrates of erwinia carotovora grown on a defined medium containing citrus pectin as the sole carbon source. the heat-labile elicitors were highly purified by cation-exchange chromatography on a cm-sephadex (c-50) column, followed by agarose-affinity chromatography on a bio-gel a-0.5m gel filtration column. the heat-labile elicitor activity co-purified with two alpha-1,4-endopo ...198416663385
changes in properties of phytopathogenic bacteria effected by plasmid prd1.transfer of plasmid prd1 from escherichia coli k 12j62 -1 to phytopathogenic bacteria, agrobacterium tumefaciens, xanthomonas beticola and erwinia carotovora subsp. carotovora caused changes in conjugant properties not determined by the plasmid and the emergence of the properties not present in the parent strains. clones have been obtained with intermediate properties between donor and recipient, including those with altered or lost virulence. in transconjugants of a. tumefaciens virulence incre ...19846375619
cloning and characterization of reca genes from proteus vulgaris, erwinia carotovora, shigella flexneri, and escherichia coli b/r.the reca genes of proteus vulgaris, erwinia carotovora, shigella flexneri and escherichia coli b/r have been isolated and introduced into escherichia coli k-12. all the heterologous genes restore resistance to killing by uv irradiation and the mutagen 4-nitroquinoline-1-oxide in reca- e. coli k-12 hosts. recombination proficiency is also restored as measured by formation of lac+ recombinants from duplicated mutant lacz genes and the ability to propagate phage lambda derivatives requiring host re ...19846090408
transposon tn5 mutagenesis in erwinia carotovora subsp. carotovora and e. carotovora subsp. atroseptica.in matings between escherichia coli 2492(pjb4ji) and erwinia carotovora subsp. carotovora ecc71 and e. carotovora subsp. atroseptica eca12, kmr gms transconjugants were obtained at high frequencies, indicating instability of the mu-containing plasmid pjb4ji and transposition of tn5 into the recipient genome. this was verified by southern blot hybridization with prz102 dna containing tn5 as the 32p-labeled probe. examination of kmr gms transconjugants of ecc71 and eca12 disclosed that a proportio ...19846321435
mutagenesis of erwinia carotovora subsp. carotovora with bacteriophage mu d1 (apr lac cts62): construction of his-lac gene fusions.the bacteriophage mu d1(apr lac cts62 ) obtained from an escherichia coli double lysogen carrying the defective mu d1 phage and a mu-p1 hybrid phage was utilized as a vector for phage mutagenesis in erwinia carotovora subsp. carotovora. among ampicillin-resistant transductants. 1.4% were auxotrophs. the synthesis of beta-galactosidase was derepressed upon starvation for histidine in two different his-lac fusion strains.19846233263
chromosomal mapping in erwinia carotovora subsp. carotovora with the incp plasmid r68::mu.conjugational gene transfer was established in erwinia carotovora subsp. carotovora scri193 by using plasmid r68::mu c+ to mobilize the chromosome into multiply mutant recipients. it was observed that although the plasmid alone mobilized markers randomly at a frequency of ca. 10(-5) chromosomal recombinants per donor, the presence of a mu prophage on the chromosome of the donor increased the frequency of mobilization of markers adjacent to the prophage by up to 10-fold. using this system it was ...19852933392
[characteristics of transduction in erwinia by phage 59].a temperate bacteriophage 59 from polylysogenic strain erwinia carotovora 268 transduces the following genetic markers: arg+, ilv+, leu+, met+, thr+, thy+, trp+, ura+. the transduction frequencies varied from 1 x 10(-8)- to 1 x 10(-6) and dependent on the multiplicity of infection, uv-irradiation of transducing bacteriophage, the nature of phage lysates. the characteristics of single transductants have been studied. analysis of the obtained results suggests bacteriophage 59 to perform the genera ...19853870351
cloning and expression in escherichia coli of pectinase genes of erwinia carotovora subsp. carotovora.genes coding for an endo-pectate lyase, an exo-pectate lyase, and an endopolygalacturonase of erwinia carotovora subsp. carotovora ecc71 were cloned in escherichia coli hb101, using the cosmid phc79. the products of the cloned pectinase genes paralleled their counterparts in strain ecc71 in isoelectric mobility, mode of substrate degradation, and ability to macerate potato tuber tissue.19853888112
cloning of the pectate lyase genes from erwinia carotovora and their expression in escherichia coli.a hybrid cosmid coding for pectate lyase (pl) activity was identified from an erwinia carotovora genomic library by an immunological screening method. a 7-kb dna fragment was identified which codes for three proteins identical in size to proteins with pl activity purified from e. carotovora culture supernatants. the three proteins had apparent mrs of 41, 44 and 44 x 10(3) as estimated by sds-page. none of the pls were exported from escherichia coli strain hb101 but all were found in the periplas ...19853896936
arab gene and nucleotide sequence of the arac gene of erwinia carotovora.the arab and arac genes of erwinia carotovora were expressed in escherichia coli and salmonella typhimurium. the arab and arac genes in e. coli, e. carotovora, and s. typhimurium were transcribed in divergent directions. in e. carotovora, the arab and arac genes were separated by 3.5 kilobase pairs, whereas in e. coli and s. typhimurium they were separated by 147 base pairs. the nucleotide sequence of the e. carotovora arac gene was determined. the predicted sequence of arac protein of e. caroto ...19853902795
efficient transformation of erwinia carotovora subsp. carotovora and e. carotovora subsp. atroseptica.we used a modified version of the method of hanahan (d. hanahan, j. mol. biol. 166:557-580, 1983) to transform erwinia carotovora subsp. carotovora and e. carotovora subsp. atroseptica with the plasmids pbr322, pbr325, and pat153. the transformation frequency ranged from 1 x 10(2) to 4 x 10(4) colonies per micrograms of plasmid dna. the nature of these transformants was confirmed by plasmid analysis. cole1-based plasmids make potentially useful cloning vectors for the study of genes involved in ...19853968041
structure of the core oligosaccharide from lipopolysaccharide of erwinia carotovora.the lipopolysaccharide of erwinia carotovora was analyzed by quantitative sugar analysis, methylation analysis, and chromic oxide oxidation. this led to the following structure of the core oligosaccharide: (formula; see text).19853972773
the virulence of clinical and environmental isolates of campylobacter jejuni.the virulence of campylobacter jejuni and c. coli isolated from various water sources was compared with that of clinical strains by in vitro assays of adhesion, invasion and cytotoxicity to hela cells. variation in degree of attachment was observed, but this did not appear to be related to strain source, however, water strains were less invasive and less cytotoxic to hela cells than clinical strains as shown by immunofluorescence and electron microscopy. these differences were particularly evide ...19853973380
reca is required in the induction of pectin lyase and carotovoricin in erwinia carotovora subsp. carotovora.pectin lyase (pnl) and the bacteriocin carotovoricin (ctv) were induced in erwinia carotovora subsp. carotovora 71 by the dna-damaging agents mitomycin c, nalidixic acid, and uv light. to determine whether the reca product was involved in the expression of these damage-inducible phenotypes, we cloned the e. carotovora subsp. carotovora reca+ gene, inactivated it by tn5 insertion, and constructed an e. carotovora subsp. carotovora reca::tn5 strain by gene replacement via homologous recombination. ...19854044526
purification of rna polymerase and transcription-termination factor rho from erwinia carotovora.erwinia carotovora rna polymerase consists of the holoenzyme structure sigma 2 beta beta' sigma as found in escherichia coli and other bacteria. e. carotovora rna polymerase can synthesize rna using lambda, t7 of t4 dna as templates; however, it is two times less active on these templates and is more temperature-sensitive than the e. coli enzyme. the alpha subunit of the e.. carotovora enzyme is lower in molecular mass than its e. coli counterpart. the sigma factors from e. coli and e. carotovor ...19852578393
an improved dna sequencing strategy.a modification of hong's systematic dna sequencing strategy is described. the original procedure has been simplified and transfectant yield increased. after dnase i limited cleavage in the presence of mn2+, the single-cut linear dna does not have to be separated from supercoiled or open circular dna on an agarose gel. after ligation, the dna is digested with a second restriction endonuclease for which a unique cleavage site resides between the insert and the first restriction endonuclease cuttin ...19852992313
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
effects of a mutation that eliminates udp glucose-pyrophosphorylase on the pathogenicity of erwinia carotovora subsp. carotovora.a nonpathogenic mutant of erwinia carotovora obtained by mu d1 mutagenesis was defective in the ability to utilize several carbon sources. the basis of the mutation was analyzed biochemically and shown to be a defect in the ability to form udp glucose-pyrophosphorylase. the nonpathogenic phenotype of the mutant was caused by its sensitivity to galactose.19852995320
evidence that polygalacturonase is a virulence determinant in erwinia carotovora.polygalacturonase (pg) was purified from erwinia carotovora ec. a hybrid cosmid, psh711, that encodes pg activity but not pectate lyase activity was identified from an e. carotovora genomic library by an immunological screening method. a cell extract of escherichia coli cells containing psh711 was able to produce plant tissue maceration when spotted on carrot, potato, or turnip slices. in addition, the e. coli strain containing this plasmid was able to macerate carrot, potato, and turnip slices. ...19852997134
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
requirement for two or more erwinia carotovora subsp. carotovora pectolytic gene products for maceration of potato tuber tissue by escherichia coli.several genes encoding enzymes capable of degrading plant cell wall components have been cloned from erwinia carotovora subsp. carotovora ec14. plasmids containing cloned ec14 dna mediate the production of endo-pectate lyases, exo-pectate lyase, endo-polygalacturonase, and cellulase(s). escherichia coli strains containing one of these plasmids or combinations of two plasmids were tested for their ability to macerate potato tuber slices. only one e. coli strain, containing two plasmids that encod ...19863013836
soluble asparaginase-dextran conjugates show increased circulatory persistence and lowered antigen reactivity.oxidized dextrans of increasing molecular weight were bound covalently to erwinia carotovora asparaginase. the resulting conjugates retained 50% of their enzyme activity and showed marked resistance to proteolysis by trypsin and chymotrypsin and inactivation by asparaginase-specific antibody. when tested in-vivo, the larger molecular weight conjugates showed prolonged circulatory survival in both immune and non-immune animals and failed to elicit full type iii hypersensitivity or anaphylactic re ...19862423675
large-scale recovery and purification of l-asparaginase from erwinia carotovora.a large-scale process was developed to purify gram quantities of a therapeutic enzyme, l-asparaginase, from submerged cultures of erwinia carotovora. cells were harvested from 150 l of fermentation broth and washed. a cellular acetone powder was prepared and extracted with ph 9.5 borate buffer. after continuous centrifugation and filtration to remove cell debris, the acetone powder extract was adjusted to ph 7.7 and adsorbed onto a 16-l cm-sepharose fast flow column, with a precolumn packed with ...19863752984
comparison of pectic enzymes produced by erwinia chrysanthemi, erwinia carotovora subsp. carotovora, and erwinia carotovora subsp. atroseptica.erwinia spp. that cause soft-rot diseases in plants produce a variety of extracellular pectic enzymes. to assess the correlation between patterns of pectic enzyme production and taxonomic classification, we compared the enzymes from representative strains. supernatants obtained from polygalacturonate-grown cultures of nine strains of erwinia chrysanthemi, three strains of e. carotovora subsp. carotovora, and three strains of e. carotovora subsp. atroseptica were concentrated and subjected to ult ...19863752996
host-pathogen interactions : xxix. oligogalacturonides released from sodium polypectate by endopolygalacturonic acid lyase are elicitors of phytoalexins in soybean.recent studies have demonstrated that an apparently homogeneous preparation of an alpha-1,4-d-endopolygalacturonic acid lyase (ec 4.2.2.2) isolated from the phytopathogenic bacterium erwinia carotovora induced phytoalexin accumulation in cotyledons of soybean (glycine max [l.] merr. cv wayne) and that this pectin-degrading enzyme released heat-stable elicitors of phytoalexins from soybean cell walls, citrus pectin, and sodium polypectate (kr davis et al. 1984 plant physiol 74: 52-60). the presen ...198616664663
antagonism of lactic acid bacteria against phytopathogenic bacteria.a variety of lactic acid bacteria, isolated from plant surfaces and plant-associated products, were found to be antagonistic to test strains of the phytopathogens xanthomonas campestris, erwinia carotovora, and pseudomonas syringae. effective "in vitro" inhibition was found both on agar plates and in broth cultures. in pot trials, treatment of bean plants with a lactobacillus plantarum strain before inoculation with p. syringae caused a significant reduction of the disease incidence.198616347150
a rapid procedure for purifying pectate lyase from erwinia carotovora based on substrate affinity.to purify pectate lyase produced by erwinia carotovora subsp. carotovora, we used the supernatant from 48-h-old cultures grown in broth containing sodium polypectate and yeast extract. the supernatant was combined with the enzyme substrates sodium polypectate and polygalacturonic acid, which were then precipitated with cacl(2). after the precipitate was washed, pectate lyase was eluted with 1.0 m nacl.198716347348
fermentation of d-xylose and l-arabinose to ethanol by erwinia chrysanthemi.erwinia spp. are gram-negative facultative anaerobes within the family enterobacteriacae which possess several desirable traits for the conversion of pentose sugars to ethanol, such as the ability to ferment a broad range of carbohydrates and the ease with which they can be genetically modified. twenty-eight strains of erwinia carotovora and e. chrysanthemi were screened for the ability to ferment d-xylose to ethanol. e. chrysanthemi b374 was chosen for further study on the basis of its superior ...198716347426
induction of defense responses in cultured parsley cells by plant cell wall fragments.cell suspension cultures of parsley (petroselinum crispum) accumulated coumarin phytoalexins and exhibited increased beta-1,3-glucanase activity when treated with either a purified alpha-1,4-d-endopolygalacturonic acid lyase from erwinia carotovora or oligogalacturonides solubilized from parsley cell walls by endopolygalacturonic acid lyase. coumarin accumulation induced by the plant cell wall elicitor was preceded by increases in the activities of phenylalanine ammonia lyase (pal), 4-coumarate: ...198716665599
the arac gene of citrobacter freundii.the arac gene of citrobacter freundii was cloned into plasmid pbr322 and expressed in escherichia coli and salmonella typhimurium. the nucleotide sequence and the predicted translational product were determined and compared to those of e. coli, s. typhimurium and erwinia carotovora. the predicted translational product is 281 amino acids (aa) long, identical in size to that of s. typhimurium, and is 11 and 29 aa shorter than that of e. coli and e. carotovora, respectively. the nucleotide sequence ...19872965663
[complementation analysis of ts mutants of the temperate phage 59 of erwinia carotovora 268]. 19873508923
[the monosaccharide composition of erwinia carotovora subsp. carotovora cells]. 19873508943
[reproduction and morphogenesis of phage e105 of erwinia carotovora 268]. 19873508944
hemagglutinating activity in phytopathogenic bacteria surface compounds.extracellular components of plant pathogenic bacteria were obtained from their culture medium as well as from the whole cells by using nacl 1 m, ph 6.0; 20% sucrose dissolved in 0.03 m tris buffer, ph 8.0; or 0.05 m na2edta. all the extracts from erwinia carotovora subsp. carotovora, xanthomonas campestris pv. campestris, pseudomonas syringae pv. phaseolicola, xanthomonas campestris pv. phaseoli, pseudomonas solanacearum, and erwinia carotovora subsp. atroseptica, were assayed for hemagglutinati ...19873625474
use of tnphoa to enrich for extracellular enzyme mutants of erwinia carotovora subspecies carotovora.soft rot erwinia species secrete a range of enzymes into the extracellular environment. therefore, the genetically amenable erwinia system is a useful model for the study of protein secretion by gram-negative bacteria. we have used a lambda-sensitive derivative of erwinia carotovora subspecies carotovora (ecc) and the transposon tnphoa, to isolate a range of extracellular enzyme mutants. the use of tnphoa provides an enrichment for extracellular enzyme mutants over other transposon-based systems ...19872834624
characterization of type 1 and mannose-resistant fimbriae of erwinia spp.type 1 fimbriae from erwinia carotovora subsp. carotovora and mannose-resistant fimbriae from erwinia rhapontici were purified and characterized. the type 1 fimbrillin had an apparent molecular weight of 16,500; that of the mannose-resistant fimbrillin was 18,000. the amino-terminal amino acid sequences of the two fimbrillins were related, but tryptic peptide maps showed significant differences between the proteins. no serological cross-reaction was found between the two fimbrial filaments, nor ...19872883172
characterization of the erwinia carotovora pelb gene and its product pectate lyase.the pelb gene encodes pectate lyase b, one of three pectate lyases identified in erwinia carotovora ec. pectate lyase b was purified from escherichia coli containing the pelb gene on a recombinant plasmid. the activity of the protein was optimal at a ph of 8.3. the amino acid composition, n-terminal amino acid sequence, and c-terminal peptide sequence were determined and compared with the polypeptide sequence deduced from the dna sequence of pelb. purified pectate lyase b started at amino acid 2 ...19873040692
a note on the microbiology of retail packs of prepared salad vegetables.retail packs of mixed, prepared salad vegetables from two different manufacturers were stored at 7 degrees c until the end of storage-life (sell-by date plus 1 d), when the microbial flora was examined. the quality of the salads was acceptable at the end of storage life. the oxygen concentrations in packs were lower, and the carbon dioxide concentrations were higher, than those in air. high numbers of bacteria were present, with pseudomonas spp. and enterobacter agglomerans predominating in pack ...19873126172
[cloning of the regulator gene of erwinia carotovora repressing pectate lyase ptla gene expression].pectate lyase synthesis in the cells of erwinia carotovora ela 49 is induced by polypectate. this suggested that the erwinia chromosomes carried a regulator gene responsible for negative regulation of the pectate lyase gene expression. in the present study the regulator gene controlling expression of one of the pectate lyase structural genes was cloned and designated as ptla gene. for this purpose a genetic system with the tester plasmid ppc624 as the main element was constructed. the tester pla ...19873307613
microbial coagulation of alfalfa green juice.of the different bacterial strains isolated from alfalfa raw material, nine were able to coagulate the protein fraction of alfalfa green juice. the two strains showing the highest efficiency were further used for coagulation experiments. they were classified as erwinia carotovora and escherichia coli. juice samples inoculated (1:10 to 1:100) with stationary-phase cultures were efficiently coagulated. the amount of protein recovered was equivalent to that obtained when the juice was heat treated. ...19873314708
methanol production from the degradation of pectin by human colonic bacteria.when ingested, pectin can lower serum cholesterol levels in humans. pectin is degraded by fecal bacteria in the colon. we examined the release of methanol (meoh) by this degradation. a 0.2% glucose (2 g/l) mixture was used as the control medium. a pure culture of pectinolytic erwinia carotovora was the control bacterium. the chief substrates were, in set 1, 0.2% pectin (2 g/l) and, in set 2, 0.1% glucose (1 g/l) and 0.1% pectin (1 g/l). cultures of fecal bacteria and e carotovora grew for 72 h i ...19883364401
characterization of the erwinia carotovora pela gene and its product pectate lyase a.the pela gene from erwinia carotovora strain ec encodes pectate lyase a (pla). the gene was cloned and sequenced. the pla was purified from escherichia coli containing the pela gene on a recombinant plasmid. the optimum ph for the enzyme reaction is at ph 8.5; the optimum temperature for the enzyme reaction is at 40 degrees c; and the purified pla alone can macerate potato slices. nucleotide sequence of pela and the deduced polypeptide sequence of pla were compared with the pelb gene of e. carot ...19883371662
microorganisms associated with mouldiness of dried yam chips and their prevention.the broad objective of this study was to isolate and identify the microorganisms causing mouldiness of stored yam chips and to look for ways of preventing the problem. microorganisms isolated included aspergillus flavus, a. glaucus, a. nidulans, a. niger, a. ochraceous, a. tamarii, a. candidus, penicillium oxalicum, trichoderma longibrachyatum, rhizopus nigricans, cylindrocarpon radicicola, neurospora crassa, botryodiplodia theobromae, bacillus subtilis, bacillus cereus, erwinia carotovora and s ...19883231261
bacteriophage t4 resistant mutants of the plant pathogen erwinia carotovora.bacterial lipopolysaccharide (lps) has been implied in a variety of pathogenic and symbiotic plant-bacterium interactions. in order to study the role of lps in pathogenicity of erwinia carotovora, a broad host range phytopathogen, we isolated lps defective mutants of two subspecies of erwinia carotovora, subsp. carotovora (ecc) and subsp. astroseptica (eca). this was accomplished by using the escherichia coli phage t4 as a selective agent. screening of erwinia isolates revealed that some of them ...19883241545
conservation of an atp-binding domain among reca proteins from proteus vulgaris, erwinia carotovora, shigella flexneri, and escherichia coli k-12 and b/r.the purified reca proteins encoded by the cloned genes from proteus vulgaris, erwinia carotovora, shigella flexneri, and escherichia coli b/r were compared with the reca protein from e. coli k-12. each of the proteins hydrolyzed atp in the presence of single-stranded dna, and each was covalently modified with the photoaffinity atp analog 8-azidoadenosine 5'-triphosphate (8n3atp). two-dimensional tryptic maps of the four heterologous reca proteins demonstrated considerable structural conservation ...19883286605
[limited proteolysis of diphtheria toxoids in the periplasm of escherichia coli and erwinia carotovora]. 19883048950
microcalorimetric determination of substrate specificity of l-asparaginase in escherichia coli and erwinia carotovora. 19883065395
[construction of plasmids coding for diphtheria toxin fragments].plasmids coding different nontoxic derivatives (toxoids) of the diphtheria toxin were constructed. a secretion of toxoids that carry a signal sequence was found in the periplasmic space of e. coli and erwinia carotovora. toxoids without a signal sequence appear in the cytoplasm. we believe that the toxoids secreted in e. coli and e. carotovora cells undergo a limited proteolysis. according to the molecular weights of the fragments there are three targets for proteolysis. one of them being just b ...19883065616
[restriction map of permuted dna of erwinia carotovora temperate bacteriophage 59].the cyclic permutation and terminal redundancy were found in the genomes of erwinia carotovora temperate bacteriophage 59 by electron microscopic studies. the headful mechanism for bacteriophage dna cleavage and packaging during the phage morphogenesis was confirmed by the restriction analysis technique. restriction map of the bacteriophage 59 dna was constructed for restriction endonucleases bamhi, bg1ii, eco31, sa1i, smai, ecori.19882895890
[restriction map of the temperate phage e105 from the polylysogenic culture of erwinia carotovora 268].dna structure of the temperate bacteriophage e105 from polylysogenic culture of erwinia carotovora 268 has been studied. the viral 29.12-29.17 md dna has been shown to be linear and nonpermuted. the complete restriction map of the viral dna has been constructed for mvai and hpai and partial for eco31 restriction endonucleases based on the pair hydrolysis of the native dna as well as its fragments. altogether, 19 sites for restriction endonucleases have been localized on bacteriophage dna.19882841593
role of antibiosis in competition of erwinia strains in potato infection courts.erwinia carotovora subsp. betavasculorum strains produced a bactericidal antibiotic in vitro that inhibited a wide spectrum of gram-negative and gram-positive bacteria. the optimum temperature for production was 24 degrees c, and the addition of glycerol to culture media enhanced antibiotic production. antibiotic production by these strains in the infection court of potato was the principal determinant enabling it to gain ascendancy over competing antibiotic-sensitive erwinia carotovora subsp. c ...198816347633
genetically engineered erwinia carotovora in aquatic microcosms: survival and effects on functional groups of indigenous bacteria.the survival of genetically engineered erwinia carotovora l-864, with a kanamycin resistance gene inserted in its chromosome, was monitored in the water and sediment of aquatic microcosms. the density of genetically engineered and wild-type e. carotovora strains declined at the same rate, falling in 32 days below the level of detection by viable counts. we examined the impact of the addition of genetically engineered and wild-type strains on indigenous bacteria belonging to specific functional g ...198916347942
structure of a pectic polysaccharide fraction from zea shoots.a pectic fraction, accounting for about 0.3% of the total cell wall polysaccharide, was derived from the hot water extract of an insoluble fraction of the buffer-homogenate of zea shoots. the pectic polysaccharide fraction was characterized by fragmentation analysis after hydrolysis with acid and erwinia carotovora pectate lyase. the results suggest that the fraction consists of mostly a linear homopolygalacturonan with neutral sugar components or a homogalacturonan and a rhamnogalacturonan with ...198916666623
cloning and characterization of a pectate lyase gene from erwinia carotovora ec153.a pel gene cloned from strain ec153 of erwinia carotovora encoded a pectate lyase that macerated plant tissue with moderate efficiency. this gene, called pel153, was sequenced and found to possess considerable homology with a pectate lyase gene from yersinia pseudotuberculosis. the yersinia protein, however, was truncated at the carboxyl terminal end relative to the erwinia gene product and had a lower isoelectric point. the erwinia pel153 gene was overexpressed in cells of escherichia coli, and ...19892520159
improvement of dna transfer frequency and transposon mutagenesis of erwinia carotovora subsp. betavasculorum.the production of antibiotics and their role in microbial competition under natural conditions can be readily studied by the use of transposon mutants. several antibiotic-producing strains of erwinia carotovora subsp. betavasculorum were unable to accept foreign dna. a plasmid delivery system was developed, using ethyl methanesulfonate mutagenesis, which entailed isolating e. carotovora subsp. betavasculorum mutants able to accept foreign dna and transfer it to other strains. this enabled transp ...19892543291
isolation and characterisation of transposon-induced mutants of erwinia carotovora subsp. atroseptica exhibiting reduced virulence.the blackleg pathogen erwinia carotovora subsp. atroseptica (eca) causes an economically important disease of potatoes. we selected a genetically amenable eca strain for the genetic analysis of virulence. tn5 mutagenesis was used to generate nine mutants which exhibited reduced virulence (rvi-) of strain scri1043. following physiological characterisation, mutants were divided into three classes: (1) auxotrophs; (2) extracellular enzyme mutants; and (3) a growth rate mutant. the isolation of thes ...19892549365
modification of l-asparaginase from erwinia carotovora using human serum albumin.l-asparaginase from erwinia carotovora was modified by coupling with human serum albumin. the complex retained 80% of the activity, shifted the ph optima to 7.0, and lowered the km value by tenfold, but the temperature optima remained at 37 degrees c. it was also found to be heat-resistant.19892604913
attenuation of asparaginase-induced hyperglycemia after substitution of the erwinia carotovora for the escherichia coli enzyme preparation.l-asparaginase, an enzyme with established antileukemic activity, increases the induction rate and duration of remission of acute lymphoblastic leukemia when added to vincristine and prednisone for induction therapy. enzymes derived from two different bacterial sources (escherichia coli and erwinia carotovora) are in common use. these enzymes may be associated with toxic reactions of differing frequency and severity. specifically, the complication of enzyme-induced hyperglycemia may be seen more ...19892643458
extracellular and periplasmic isoenzymes of pectate lyase from erwinia carotovora subspecies carotovora belong to different gene families.pectate lyase (pel) plays a crucial role in the maceration of vegetables by soft rot erwinia spp. we have characterized the four pel isoenzymes of erwinia carotovora subspecies carotovora strain scri193. in this paper we concentrate on two isoenzymes which have different locations in scri193: plb is periplasmic and plc is extracellular. comparison of the gene products and nucleotide sequences of pelb and pelc allowed us to assign them to different gene families. in addition, we have identified a ...19892695748
[an electron microscopic study of the structure of erwinia carotovora 268 bacteria].the electron-microscopic studies have shown that the bacterium erwinia carotovora 268 (a producer of l-asparaginase) does not differ essentially from other representatives of enterobacteria. when growing on the artificial nutrient medium m9, the division of bacteria is inhibited and the bacterial cell itself elongates. certain functionally specified structural peculiarities such as the formation of the protein microcapsule, formation of cavities and compacted sites of the cytoplasm and mucus on ...19892695793
[characteristics of the lipopolysaccharides in erwinia carotovora subsp. carotovora 8883 isolated by various methods]. 19892725355
characterization of a monoclonal antibody against active pectate lyase from erwinia carotovora.a monoclonal antibody (2e2) produced against pectate lyase from erwinia carotovora ssp. carotovora reacted with a 41- and a 44-kilodaltion protein on western blots of concentrated erwinia culture supernatants resolved by sodium dodecyl sulfate - polyacrylamide gel electrophoresis. it was unequivocally shown that monoclonal 2e2 reacted with an active form of pectate lyase by affinity purifying the antigen with the monoclonal. the affinity-purified antigen was enzymatically active and moved as a s ...19892766119
two haemophilus influenzae rd genes that complement the reca-like mutation rec-1.two haemophilus influenzae rd genes each complemented the pleiotropic defects of the reca-like mutation rec-1. one gene, fec, was isolated on a 3.6-kilobase-pair ecori restriction fragment by complementation of the fec- phenotype of bacteriophage lambda. the other gene, rec, was identified on a 3.1-kilobase-pair ecori fragment by southern hybridization by using reca-like gene probes from erwinia carotovora and pseudomonas aeruginosa pao. in a rec-1 strain of h. influenzae, the cloned genes resto ...19892785104
l-asparaginase from erwinia carotovora. an improved recovery and purification process using affinity chromatography.a large-scale process was developed to purify l-asparaginase from submerged cultures of erwinia carotovora. cells from 880 l of fermentation broth were harvested and washed using a plate and frame type filter press. a cellular acetone powder was prepared from the washed cells by suspending the cells twice in acetone and the residual acetone was removed by washing the acetone powder in the filter press with 10 mm phosphate buffer (ph 7.0). the cellular acetone powder was extracted with 10 mm bora ...19892802597
enumerating low densities of genetically engineered erwinia carotovora in soil.an inexpensive, quantitative, and sensitive technique was developed for detection of genetically engineered erwinia carotovora in soil samples. enrichment media, antibiotic resistance, and most probable number (mpn) analysis were used to enumerate as few as 1 to 10 target cells/10 g soil. the mpn technique recovered significantly higher cell densities than plating; however, densities estimated by the two techniques were strongly correlated. after inoculation of soil microcosms with genetically e ...19901366379
facile isolation of endo-pectate lyase from erwinia carotovora based on electrostatic interaction.endo-pectate lyase (pate) from erwinia carotovora was selectively cosedimented with extracellularly produced lipopolysaccharide-lipid complex (lpslc) through dialysis of the cell free culture broth. the selective isolation of pate was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. the cosedimentation of the pate with lpslc was initiated by decreasing conductivity of the solution and terminated at approx 1 m siemens (mscm-1). as much as 62% of pate activity in the culture ...19902091529
[polyamines in representative taxa of coryneform and other bacteria].the composition of polyamines is studied for the first time in representatives of the genus micrococcus and taxon "conglomeratus", strains staphylococcus aureus ccm 209, deinococcus erythromyxa ccm 706 as well as of erwinia carotovora atcc 15713 polyamines, which are not extracted by perchloric acid. considerable amounts of spermine and rarely of spermidine are revealed in cells of gram positive microorganisms, that differs them from gram negative bacteria possessing high concentrations of putre ...19902115965
cdna cloning of carrot extracellular beta-fructosidase and its expression in response to wounding and bacterial infection.we isolated a full-length cdna for apoplastic (extracellular or cell wall-bound) beta-fructosidase (invertase), determined its nucleotide sequence, and used it as a probe to measure changes in mrna as a result of wounding of carrot storage roots and infection of carrot plants with the bacterial pathogen erwinia carotovora. the derived amino acid sequence of extracellular beta-fructosidase shows that it is a basic protein (pl 9.9) with a signal sequence for entry into the endoplasmic reticulum an ...19902152110
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
[erwinia carotovora as a recipient system for the natural hybrid plasmid rp4::mu cts62].the plasmid rp4::mu cts62 is transferred from escherichia coli cells into a recipient strain erwinia carotovora 268 by conjugation with the frequency 1.5-5 x 10(-7) per donor cell. the maximal frequencies of transfer are obtained by cultivation of donor and recipient cells for 3-5 h on the filters. structural and functional validity of the plasmid in transconjugants is expressed in preservation of all antibiotic-resistant markers of rp4, thermosensitivity to growth at 42 degrees c as well as spo ...19902175013
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