Publications
| Title | Abstract | Year Filter | PMID(sorted descending) Filter |
|---|
| efficacy of two therapy regimens for treatment of experimentally induced escherichia coli mastitis in cows. | the objective of the study was to monitor the effect of two therapy regimens on experimental escherichia coli mastitis. single udder quarters of 12 cows that were at least 30 d postpartum were inoculated with 1500 cfu of e. coli. the inoculation was repeated in the contralateral quarter after a 3- to 4-wk interval. initially, half of the cows were treated with antimicrobials, and the remaining half were left untreated. at the second inoculation, the cows that were originally treated were not tre ... | 1994 | 8182170 |
| recombinant gst-human osteopontin fusion protein is functional in rgd-dependent cell adhesion. | osteopontin (opn) is a secreted phosphoprotein expressed by many tumor cells, as well as a limited set of normal cells. native opn has been shown to support cell adhesion in an rgd-peptide-inhibitable fashion. here we expressed human opn in e. coli as a recombinant fusion protein with glutathione-s-transferase (gst). we report that the gst-opn fusion protein has functional activity. pap2 (ras-transformed, metastatic murine nih 3t3) and mda-mb-435 human mammary carcinoma cells bound to gst-opn in ... | 1994 | 8175899 |
| [effect of platelet-derived growth factor pdgf on replication of cultivated bovine lens epithelial cells]. | it has previously been reported that pdgf isoforms ab and bb induce an increase of cytosolic free calcium in cultured bovine lens epithelial cells in a dose-dependent manner. to evaluate the biological capacity of pdgf, we investigated the proliferative response of bovine lens epithelial cells to stimulation with pdgf-aa, -ab and -bb. since various unspecific components of serum-containing media act as mitogenes and mask the effect of pdgf, serum-free culture conditions were a prerequisite for g ... | 1994 | 8173260 |
| development of a novel type of cloning vector for suicide selection of recombinants. | based on the primary structure of the rat corticostatin r4 (antimicrobial defensin ratnp-1), a synthetic gene was designed, synthesized, and inserted into the iptg-inducible prokaryotic expression vector pflag, with an additional 13 codons between the flag sequence and the synthetic r4 sequence. this construct, n-p1.2, was further developed by inclusion of multiple cloning sites right after the flag sequence, forming a new plasmid pscv-1. escherichia coli transformants containing pscv-1 or n-p1. ... | 1994 | 8172659 |
| refolding and characterization of human recombinant heparin-binding neurite-promoting factor. | heparin-binding neurite-promoting factor (hbnf) is a highly basic, cysteine-rich 136-residue protein, and a member of a new class of heparin-binding proteins. it exhibits a neurite-outgrowth promoting activity and its expression is both temporally and spacially regulated during fetal and postnatal development. a high interspecies sequence conservation suggests important, presently unknown, biological functions. hbnf is structurally and most likely functionally related to the product of a develop ... | 1994 | 8167469 |
| characterization of the saccharomyces cerevisiae nuclear gene cyb3 encoding a cytochrome b polypeptide of respiratory complex ii. | computer-assisted structural analysis of the predicted product of the previously described open reading frame (orf) ykl4 located on the left arm of chromosome xi of saccharomyces cerevisiae revealed a high degree of similarity (> 50%) to bovine cytochrome b560, the sdhc polypeptide of the escherichia coli succinate dehydrogenase (sdh) complex and the protein specified by orf137 located on the chloroplast dna of marchantia polymorpha. disruption of the yeast gene severely impaired mitochondrial f ... | 1994 | 8152421 |
| effects of recombinant human cytokines on mitogen-induced bovine peripheral blood mononuclear cell proliferation. | the effects of interleukin 1 alpha (il-1 alpha), il-1 beta, il-2, il-3, il-4, il-5, il-6, il-7, il-8, and insulin growth factor-1 (igf-1) on proliferation of mitogen-stimulated bovine peripheral blood mononuclear cells (pbmc) were determined. four concentrations of each cytokine were incubated 48 or 96 h with pbmc alone or with pbmc and three concentrations of three different mitogens. the mitogens included concanavalin a (con a) phytohemagglutinin (pha), and escherichia coli 055:b5 lipopolysacc ... | 1993 | 8142606 |
| the extracellular nuclease of serratia marcescens: studies on the activity in vitro and effect on transforming dna in a groundwater aquifer microcosm. | a quantitative endonuclease assay, which relies on the introduction of single and double strand breaks into supercoiled plasmid dna, was used to study the activity of the extracellular nuclease of serratia marcescens sm6 in buffer and in groundwater. the parallel enzyme concentration-dependent production of relaxed and linear plasmid molecules suggests that the nuclease produces single and double strand breaks in duplex dna. bovine serum albumin stimulated the nuclease activity towards dna and r ... | 1994 | 8141644 |
| differential expression of bovine adseverin in adrenal gland revealed by in situ hybridization. cloning of a cdna for adseverin. | adseverin is a ca(2+)-dependent actin filament-severing protein that is presumed to have a regulatory function in exocytosis by affecting the organization of the microfilament network underneath the plasma membrane. as the first step to investigate whether adseverin has the expected function in vivo, we have cloned a cdna for bovine adseverin. analysis of the nucleotide sequence revealed the following points. 1) adseverin consisted of 715 amino acid residues, and its predicted molecular mass was ... | 1994 | 8119933 |
| recombinant bovine somatotropin blunts plasma tumor necrosis factor-alpha, cortisol, and thromboxane-b2 responses to endotoxin in vivo. | in vivo studies determined the effects of recombinant bovine somatotropin (bst; sometribove) administration (0.1 mg/kg.day, im) to calves on the increases in plasma immunoreactive tumor necrosis factor-alpha (tnf alpha), prostacyclin [6-keto-prostaglandin f1 alpha (6kp)], thromboxane-b2 (txb), and cortisol (c) that occurred after endotoxin challenge (et). two ets were administered 5 days apart to test the effect of bst on the natural attenuation of hormone and cytokine responses that occurs afte ... | 1994 | 8119146 |
| isolation and characterization of endonuclease viii from escherichia coli. | endonuclease viii, a novel presumptive dna repair enzyme, was isolated from escherichia coli by fplc1 purification. the enzyme was found in strains that contained or lacked endonuclease iii and was purified by radial flow s-sepharose, mono s, phenyl-superose, and superose 12 fplc. examination of the properties of endonuclease viii showed it to have many similarities to endonuclease iii. dna containing thymine glycol, dihydrothymine, beta-ureidoisobutyric acid, urea residues, or ap sites was inci ... | 1994 | 8110759 |
| resolution of recombination intermediates by a mammalian activity functionally analogous to escherichia coli ruvc resolvase. | a mammalian endonuclease that resolves holliday junctions has been partially purified from extracts of calf thymus and chinese hamster ovary cells. the activity acts upon (i) synthetic holliday junctions and (ii) recombination intermediates made by the escherichia coli reca protein and appears to be functionally analogous to the e. coli ruvc protein. cleavage occurs by the introduction of symmetrically related nicks in strands of like polarity to produce nicked duplex dna products. the nicks can ... | 1994 | 8106502 |
| preparation of the extracellular domain of the rabbit prolactin receptor expressed in escherichia coli and its interaction with lactogenic hormones. | the cdna of the extracellular domain of the rabbit prolactin receptor (rbprlr-ecd) was cloned in the prokaryotic expression vector ptrc99a to enable its expression in escherichia coli after induction with isopropyl-1-thio-beta-d-galactopyranoside. the bacterially expressed rbprlr-ecd protein, contained within the refractile body pellet, was solubilized in 4.5 m urea, refolded, and purified on a q-sepharose column by stepwise elution with nacl. the bioactive monomeric fraction was eluted in 0.05 ... | 1994 | 8106371 |
| complementary dna sequence of bovine cpn10 (hsp10), a chaperone protein from mitochondria. | a cdna sequence encoding a chaperone protein from bovine heart mitochondria is described. it is deduced from overlapping partial cdnas amplified in polymerase chain reactions from bovine heart cdna, using in the first instance mixtures of synthetic oligonucleotide primers based on the incomplete sequence of the cpn10 (hsp10) chaperone protein from rat liver mitochondria. the encoded bovine protein sequence is 101 amino acids in length. by analogy with the rat homologue it is likely that the init ... | 1993 | 8104547 |
| structure and copy number analyses of pap-, sfa-, and afa-related gene clusters in f165-positive bovine and porcine escherichia coli isolates. | pathogenic f165-positive escherichia coli isolates of porcine and bovine origin possess gene clusters related to extraintestinal e. coli fimbrial operons pap, sfa, and afai. probes from different segments of the pap, sfa, and afai operons were used in southern hybridization to analyze 18 f165-positive, mannose-resistant hemagglutinating e. coli isolates possessing pap- and sfa-, pap- and afa-, or pap-related sequences. only single copies of the pap-, sfa-, or afa-related sequences were found amo ... | 1993 | 8099065 |
| oxidation, cross-linking, and insolubilization of recombinant tropoelastin by purified lysyl oxidase. | the use of recombinant human tropoelastin (rte) and selected variants thereof as substrates for the assay of lysyl oxidase activity in vitro was explored. the possibility was also assessed that an insoluble elastin-like product could be generated from this elastin precursor in the absence of other macromolecules found associated with elastin in vivo. rte was more efficiently oxidized by lysyl oxidase than the insoluble chick aorta elastin substrate conventionally used. anionic amphiphilic elasti ... | 1993 | 8098038 |
| chaperonins dependent increase of cu,zn superoxide dismutase production in escherichia coli. | over-expression of the chaperonins groel and groes significantly suppressed the temperature-dependent pattern of expression of cu,zn superoxide dismutases in escherichia coli and increased the yield of active enzyme. the results obtained indicate that chaperonins prevent degradation of metal-deficient enzyme molecules. groel was shown to form a complex with unfolded cu,zn superoxide dismutase in vitro, confirming that groel can interact with beta-stranded proteins. | 1993 | 8097729 |
| a modified form of the cytochrome p450scc precursor: a new approach in studies on protein import into mitochondria. | recombinant dna was constructed providing hypersynthesis of a hybrid protein with a mrgsh6gir sequence preceding the nh2-terminus of the bovine cytochrome p450scc precursor (6his-pp450scc) in escherichia coli cells. a large-scale procedure for isolation and purification of this protein was elaborated. 6his-pp450scc was imported into isolated rat liver mitochondria and processed to the mature-sized form. as a similar procedure can be applied to other proteins the results of this work offer new op ... | 1994 | 8093069 |
| interaction of the small interstitial proteoglycans biglycan, decorin and fibromodulin with transforming growth factor beta. | we have analysed the interactions of three proteoglycans of the decorin family, decorin, biglycan and fibromodulin, with transforming growth factor beta (tgf-beta). the proteoglycan core proteins, expressed from human cdnas as fusion proteins with escherichia coli maltose-binding protein, each bound tgf-beta 1. they showed only negligible binding to several other growth factors. intact decorin, biglycan and fibromodulin isolated from bovine tissues competed with the fusion proteins for the tgf-b ... | 1994 | 8093006 |
| purification, characterization, and localization of subunit interaction area of recombinant mouse ribonucleotide reductase r1 subunit. | mammalian ribonucleotide reductase is a heterotetramer formed by the two non-identical homodimers proteins r1 and r2. we have succeeded in expressing the 90-kda mouse r1 protein in escherichia coli in an active, soluble form using the t7 rna polymerase pet vector system. to avoid inclusion bodies, the bacteria were grown at 15 degrees c with minimal concentration of the inducer isopropyl-1-thio-beta-d-galactopyranoside. after a rapid purification procedure, approximately 20 mg of pure r1 protein ... | 1994 | 8083221 |
| two-year study of endemic enteric pathogens associated with acute diarrhea in new caledonia. | a longitudinal study of diarrheal disease among patients of all ages with acute diarrhea was carried out in new caledonia from january 1990 to december 1991. stool samples from 2,088 diarrheal patients were examined for parasites, rotavirus, and bacterial pathogens. potential sources of contamination (drinking water, seawater and bovine and porcine feces) were investigated. one or more enteric pathogens were identified in 41.8 and 40.6% of the persons with diarrhea, in 1990 and 1991, respectivel ... | 1994 | 8077399 |
| a single arabidopsis gf14 isoform possesses biochemical characteristics of diverse 14-3-3 homologues. | arabidopsis cdna clones of gf14 proteins originally were isolated on the basis of their association with the g-box dna/protein complex by a monoclonal antibody screening approach. gf14 proteins are homologous to the 14-3-3 family of mammalian proteins. here we demonstrate that recombinant gf14 omega, one member of the arabidopsis gf14 protein family, is a dimeric protein that possesses many of the attributes of diverse mammalian 14-3-3 homologues. gf14 omega activates rat brain tryptophan hydrox ... | 1994 | 8061318 |
| expression of the carbohydrate recognition domain of lung surfactant protein d and demonstration of its binding to lipopolysaccharides of gram-negative bacteria. | surfactant protein d is a collagenous c-type lectin (collectin) that is found almost exclusively in the lung. a recombinant protein, composed of the neck-region and the carbohydrate binding domain of bovine lung surfactant protein d, has been overexpressed in e. coli. the recombinant protein showed the same sugar binding specificity as the native protein and was able to bind to the lipopolysaccharides of several strains of gram-negative bacteria, such as klebsiella pneumoniae, pseudomonas aerugi ... | 1994 | 8060356 |
| peptidase activity of escherichia coli aminopeptidase a is not required for its role in xer site-specific recombination. | xer site-specific recombination is required for the stable inheritance of multicopy plasmids and the normal segregation of the bacterial chromosome in escherichia coli. two related recombinases and two accessory proteins are essential for xer-mediated recombination at cer, a recombination site in the plasmid cole1. the accessory proteins, argr and pepa, function in ensuring that the xer recombination reaction acts exclusively intramolecularly, converting plasmid dimers into monomers and not vice ... | 1994 | 8057849 |
| different peptide binding specificities of hsp70 family members. | a set of heptapeptides was used to compare the relative peptide affinities of three proteins of the hsp70 family: bacterial dnak, mammalian cytosolic hsc70, and bip from mammalian er. each hsp displays a characteristic pattern of relative affinities. dnak and hsc70 are more similar to each other than to bip. a difference in peptide binding specificity may be an important determinant in adjusting an hsp70 family member to its particular cellular function. | 1994 | 8057353 |
| [expression of hybrid genes containing sequences coding for human adrenocorticotropic hormone in escherichia coli strains]. | e. coli strains producing a hybrid protein containing human adrenocorticotropic hormone (acth) and protein a of s. aureus were obtained. the sequence coding for actg was obtained from the bovine one using oligonucleotide-directed mutagenesis. the actg gene was linked with the protein a gene and its derivatives by synthetic adaptors. it was shown that each of the constructed plasmids directed the synthesis of hybrid protein in e. coli. this protein was purified on igg-sepharose. then acth was obt ... | 1994 | 8052250 |
| clonal relationships among escherichia coli serogroup o78 isolates from human and animal infections. | we investigated the clonal relationships among 63 escherichia coli strains of antigen serogroup o78 isolated from infections in humans, cattle, sheep, pigs, and chickens. both septicemic and enterotoxigenic isolates were included in the study. a main group of 55 e. coli strains consisting of 52 septicemic isolates and 3 human enterotoxigenic e. coli isolates were clustered in related clones. the remaining eight strains, four human and four animal isolates, were clonally heterogeneous. the main g ... | 1994 | 8051245 |
| molecular cloning and characterization of yeast rho gdp dissociation inhibitor. | we have previously isolated rho gdp dissociation inhibitor (rho dgi) from bovine brain and characterized it. bovine rho gdi is a protein of a m(r) of 23,421 with 204 amino acids. rho gdi inhibits the gdp/gtp exchange reaction of post-translationally lipid-modified small gtp-binding proteins (g proteins) of the rho family, including the rho, rac, and cdc42 subfamilies, and keeps them in the gdp-bound inactive form. in the present study, we first purified rho gdi from the cytosol fraction of the y ... | 1994 | 8051050 |
| characterization of mouse phosphatidylinositol transfer protein expressed in escherichia coli. | the cdna encoding mouse phosphatidylinositol transfer protein (pi-tp) was isolated by means of reverse transcriptase polymerase chain reaction. the nucleotide sequence of this cdna has a high similarity (98%) with that of rat pi-tp; the predicted amino acid sequence is 99.6% identical to that of rat pi-tp. the cdna encoding mouse pi-tp was cloned into the expression vector pet3d and the escherichia coli strain bl21(de3) was transformed with the resulting plasmid. after induction of the bacteria ... | 1994 | 8049244 |
| the high-level expression in escherichia coli of the membrane-bound form of human and rat cytochrome b5 and studies on their mechanism of function. | a t7 expression system is described for the high-level production in escherichia coli of the membrane-bound form of human and rat cytochrome b5. the cdnas of b5 have been engineered to contain a coding sequence for a four-member histidine domain at the amino-terminus of the recombinant protein permitting the use of a nickel-chelate affinity column for rapid purification of the detergent-solubilized hemoprotein. results are presented demonstrating the ability of the purified recombinant b5 protei ... | 1994 | 8037471 |
| emerging infectious diseases in the united states, 1993. | three outbreaks of disease in the united states in 1993 caused by escherichia coli o157:h7, cryptosporidium organisms, and a previously unrecognized hantavirus clearly illustrate the increasing challenges posed by emerging infectious diseases. the largest us outbreak of e. coli o157:h7 infection reported occurred as a result of contaminated hamburgers served at a fast-food restaurant chain. the largest recorded waterborne disease outbreak in us history was due to contamination of a municipal wat ... | 1994 | 8035009 |
| infrared linear dichroism reveals that a-, b-, and c-dnas in films have bases highly inclined from perpendicular to the helix axis. | infrared linear dichroism has been employed to investigate the inclination of the bases in films of poly[d(ac)].poly[d(gt)], poly[d(ag)].poly[d(ct)], and natural dnas (from escherichia coli and calf thymus). all dnas investigated assume the b-form at high (> 94%) relative humidity. poly[d(ac)].poly[d(gt)], e. coli dna, and calf thymus dna assume the a-form at low (75%) relative humidity, whereas poly[d(ag)].poly[d(ct)] assumes the c-form at low (66%) relative humidity. infrared linear dichroism ... | 1994 | 8031766 |
| domain structure of a mammalian myosin i beta. | we have determined the primary structure of a myosin i (called mammalian myosin i beta, mmi beta) from bovine brain and identified its functional domains. the protein was previously purified from brain and adrenal gland. several constructs were generated and expressed in escherichia coli as glutathione s-transferase fusion proteins and the recombinant proteins were recognized by monoclonal antibodies that recognize either "head" or "tail" domains of native myosin i. a gel overlay method was used ... | 1994 | 8022785 |
| the 86-kda subunit of autoantigen ku is a somatostatin receptor regulating protein phosphatase-2a activity. | we previously reported the immunopurification of a somatostatin receptor from the human tumoral gastric cell hgt1 using the monoclonal antibody 30f3 (reyl-desmars, f., le roux, s., linard, c., benkouka, f., and lewin, m. j. m. (1989) j. biol. chem. 264, 18789-18795). screening of a lambda gt11 hgt1-cdna library with 30f3 led us to isolate a cdna encoding an 86-kda polypeptide displaying 100% structural identity with the 86-kda subunit (p86-ku) of the ku autoantigen. recombinant p86 expressed in ... | 1994 | 8021251 |
| animals as a source of escherichia coli pathogenic for human beings. | symptoms of slt e coli-induced enteric disease in human beings include watery diarrhea, hemorrhagic diarrhea, and, in some cases, hus. the most frequent serotype associated with hus is o157:h7, although several other serotypes have also been implicated. these organisms produce slt-i, slt-ii, or both toxins. factors other than slt are implicated as virulence attributes, such as adhesins and enterohemolysins, but roles for these factors in the pathogenicity of these organisms have not been defined ... | 1994 | 8014086 |
| prediction and site-specific mutagenesis of residues in transmembrane alpha-helices of proton-pumping nicotinamide nucleotide transhydrogenases from escherichia coli and bovine heart mitochondria. | nicotinamide nucleotide transhydrogenase from bovine heart consists of a single polypeptide of 109 kd. the complete gene for this transhydrogenase was constructed, and the protein primary structure was determined from the cdna. as compared to the previously published sequences of partially overlapping clones, three residues differed: ala591 (previously phe), val777 (previously glu), and ala782 (previously arg). the escherichia coli transhydrogenase consists of an alpha subunit of 52 kd and a bet ... | 1994 | 8011636 |
| m1 protein and protein h: iggfc- and albumin-binding streptococcal surface proteins encoded by adjacent genes. | m1 protein and protein h are surface proteins simultaneously present at the surface of certain strains of streptococcus pyogenes, important pathogenic bacteria in humans. the present study concerns the structure, protein-binding properties and relationship between these two molecules. the gene encoding m1 protein (emm1) was found immediately upstream of the protein h gene (sph). both genes were preceded by a promoter region. comparison of the sequences revealed a high degree of similarity in the ... | 1994 | 8010973 |
| sequence heterogeneity of the eae gene and detection of verotoxin-producing escherichia coli using serotype-specific primers. | the distribution of the escherichia coli attaching and effacing (eae) gene in strains of verotoxin-producing e. coli (vtec) isolated from cattle and humans was studied. the majority of strains isolated from humans with bloody diarrhoea or hus and cattle with severe diarrhoea were eae positive (82 and 83% respectively). in contrast, 59% of vtec isolated from asymptomatic cattle were eae negative and of the remaining 41% that were eae positive, the majority were serotype o157. h7. the nucleotide s ... | 1994 | 8005211 |
| nadph binding and control of catalase compound ii formation: comparison of bovine, yeast, and escherichia coli enzymes. | 1. nadph binds to bovine catalase and to yeast catalases a and t, but not to escherichia coli catalase hpii. the association was demonstrated using chromatography and fluorimetry. bound nadph fluoresces in a similar way to nadph in solution. 2. bound nadph protects bovine and yeast catalases against forming inactive peroxide compound ii either via endogenous reductant action or by ferrocyanide reduction during catalytic activity in the presence of slowly generated peroxide. 3. bound nadph reduce ... | 1994 | 8002960 |
| molecular cloning, sequence analysis and expression of human pituitary glutaminyl cyclase. | a cdna clone for glutaminyl cyclase was isolated from a human pituitary cdna library and the complete dna sequence determined. the cdna clone had 1573 bp and contained an open reading frame of 1086 bases, coding for a protein of 361 amino acids and molecular mass of 40,876 da. the predicted amino acid sequence of the human cdna showed 86% sequence identity to the previously reported bovine glutaminyl cyclase sequence. a comparison of the amino acid sequences derived from the human and bovine cdn ... | 1994 | 7999256 |
| functional properties of beta(na1)val-deleted,(na2)his-->met hemoglobin synthesized in escherichia coli. | bovine hb (hemoglobin) has a low oxygen affinity in the absence of chloride ions and dpg. because of the increasing interest of this hb as a potential blood substitute we have engineered a human hb mutant with the aim of mimicking the functional properties of bovine hb. this was achieved by deleting residue beta na1 val and substituting a methionine for histidine at the beta na2 position as previously suggested by perutz and imai in 1980. our results show that the artificial mutant exhibits some ... | 1994 | 7994396 |
| phospholipase a2 engineering. structural and functional roles of the highly conserved active site residue aspartate-49. | site-directed mutagenesis and high-resolution two-dimensional (2d) proton nuclear magnetic resonance (nmr) were used to probe the structural and functional roles of a highly conserved residue, asp-49, in the interfacial catalysis by bovine pancreatic phospholipase a2 (pla2, overexpressed in escherichia coli). according to crystal structures, the side chain carboxylate of asp-49, along with the carbonyl oxygens of tyr-28, gly-30, and gly-32, and two water molecules, provides the necessary ligands ... | 1994 | 7993900 |
| baculovirus expression of bovine cytochrome p450c17 in sf9 cells and comparison with expression in yeast, mammalian cells, and e. coli. | expression of bovine 17 alpha-hydroxylase cytochrome p450 (p450c17) in insect sf9 cells using baculovirus is the fourth heterologous expression system developed for studying this enzyme. the enzyme in sf9 cell membranes has all the expected activities and closely resembles that from the other systems: cos1 cells, yeast, and escherichia coli. one curious feature of baculovirus expression of this hemoprotein is that the immunodetectable p450c17 is present in an insoluble pellet when produced in th ... | 1994 | 7986097 |
| a mechanism of proton translocation by f1f0 atp synthases suggested by double mutants of the a subunit. | three amino acid residues in the a subunit of the escherichia coli f1f0 atp synthase are essential for proton translocation: arg210, glu219, and his245. in this study, the essential glutamic acid has been relocated to position 252 with retention of function. it had been known that gln252 can be replaced by glu without significant effect. to test whether q252e would function in the absence of glu219, a "site-directed second-site suppressor" experiment was designed. saturation mutagenesis was appl ... | 1994 | 7982950 |
| enhancement of serine-sensitivity by a gene encoding rhodanese-like protein in escherichia coli. | when cells of escherichia coli are grown on lactate (or other carbon sources), an addition of serine to the medium causes growth inhibition. this growth inhibition is caused by inhibition by serine of homoserine dehydrogenase i, which is involved in threonine-isoleucine biosynthesis [hama, h., sumita, y., kakutani, y., tsuda, m., & tsuchiya, t. (1990) biochem. biophys. res. commun. 168, 1211-1216]. we have cloned and sequenced genes which enhance the serine-sensitivity. two open reading frames w ... | 1994 | 7982894 |
| smggds stabilizes nucleotide-bound and -free forms of the rac1 gtp-binding protein and stimulates gtp/gdp exchange through a substituted enzyme mechanism. | the rac proteins, rac1 and rac2, are essential components of the nadph oxidase system of phagocytes and regulate the actin assembly associated with membrane ruffling. these functions are controlled by the gtp-bound form of rac. the biochemical interaction between rac and its only known gdp-dissociation stimulator (termed smggds) was characterized. smggds was able to stimulate the incorporation of guanosine 5'-[gamma-thio]-triphosphate gtp[gamma s] into the rhoa, rac2, rac1, rap1a and cdc42hs gtp ... | 1994 | 7980444 |
| binding characteristics of s fimbriated escherichia coli to isolated brain microvascular endothelial cells. | to assess the role of s fimbriae in the pathogenesis of escherichia coli meningitis, transformants of e. coli strains with or without s fimbriae plasmid were compared for their binding to microvessel endothelial cells isolated from bovine brain cortices (bmec). the bmec's displayed a cobblestone appearance, were positive for factor viii, carbonic anhydrase iv, took up fluorescent-labeled acetylated low density lipoprotein, and exhibited gamma glutamyl transpeptidase activity. binding of s fimbri ... | 1994 | 7977653 |
| [therapy of clinically apparent forms of mastitis in lactating dairy cows using intra-mammary sulphamycin (polfa, pr)]. | sulphamycin (polfa, poland) is an intramammary preparation for the treatment of clinically apparent forms of mastitis in dairy cows. its effects were observed in 141 dairy cows treated according to the manufacturer's instructions. the animals were selected after clinical examination of the mammary gland and bacteriological examination of udder secreta. in 37 subclinically diseased cows the efficiency of treatment depended on the infectious agent and amounted to 85.2, 77.8, 73.3 and 60.0%, strept ... | 1994 | 7975049 |
| [occurrence of antibiotic resistance in bacteria causing mastitis in dairy cows]. | in 1986-1992 the susceptibility to antibiotics was investigated in 799 strains of causative agents of bovine mastitis. of these, streptococci, staphylococcus aureus, coagulase-negative staphylococci and other bacteria presented 43.5%, 22.2%, 17.9% and 16.6%, respectively. the strains examined were obtained from herds in which mastitis had been treated. within the period of investigation, the resistance of s. aureus to antibiotics reached the following values: penicillin (pnc) 33.0-20.3%, chloram ... | 1994 | 7975048 |
| isolation and characterization of the cdna encoding the channel catfish (ictalurus punctatus) form of cytochrome p450arom. | cytochrome p450arom (aromatase) is responsible for the conversion of androgens to estrogens. a cdna library was constructed from poly(a)-enriched mrna isolated from channel catfish (ictalurus punctatus) ovary and ligated into ecori-cut lambda arms. the amplified library was screened using a human aromatase dna probe. the longest clone isolated (2.1 kb) contained 20 bp of the 5'-untranslated region, a 1572-bp open reading frame, and a 509-bp 3'-untranslated region. northern blot analysis indicate ... | 1994 | 7958747 |
| the fpg protein, a dna repair enzyme, is inhibited by the biomediator nitric oxide in vitro and in vivo. | nitric oxide has been shown to be a mediator molecule in the regulation of many physiological functions. however, this small diatomic molecule in the presence of o2 generates reactive intermediates which modify dna bases and inactive enzymes at high concentrations (100 microm). we report that no generated by 1,1-diethyl-2-hydroxy-2-nitrosohydrazine (dea/no, et2nn(o)no-na+), a compound known to release no in a predictable manner, caused irreversible damage at physiological concentrations to the z ... | 1994 | 7955043 |
| a phosphatidylinositol 3-kinase is induced during soybean nodule organogenesis and is associated with membrane proliferation. | phosphatidylinositol 3-kinase (pi3k) is an important component of various receptor tyrosine kinase complexes in mammalian cells and a key enzyme required for cell division and vacuolar protein sorting in yeast. to our knowledge, this enzyme has not been characterized in plants. we report the cloning and characterization of soybean pi3k cdnas and present evidence for the induction of a distinctive form of this enzyme specific to nodule organogenesis. expression of the root form of pi3k is repress ... | 1994 | 7937816 |
| putative adhesins of anaplasma marginale: major surface polypeptides 1a and 1b. | genes for the msp1a and msp1b subunits of the anaplasma marginale surface antigen complex msp1 were previously cloned and expressed in escherichia coli. we report here the localization of msp1a and msp1b polypeptides on the surface of recombinant e. coli by using a live cell indirect immunofluorescent antibody assay. recombinant e. coli cells expressing the msp1 alpha gene or the msp1 beta gene encoding the msp1a and msp1b polypeptide subunits, respectively, were shown by a culture recovery adhe ... | 1994 | 7927726 |
| expression of recombinant bovine gamma b-, gamma c- and gamma d-crystallins and correlation with native proteins. | despite the use of bovine gamma-crystallins in numerous biophysical and chemical studies, characterization of these proteins at the molecular level is incomplete. bovine lenses have at least six gamma-crystallin protein fractions currently assigned as gamma s/i, gamma a/ivb, gamma b/ii, gamma c/iiib, gamma d/iiia and gamma e/iva. a lack of primary sequence data for corresponding gamma-crystallin genes and proteins, however, has made these assignments tenuous. to clarify these assignments, we hav ... | 1994 | 7925695 |
| the prevalence of escherichia coli o157.h7 in dairy and beef cattle in washington state. | escherichia coli o157.h7 was found in 10 of 3570 (0.28%) faecal samples from dairy cattle in 5 of 60 herds (8.3%). several tentative associations with manure handling and feeding management practices on dairy farms were identified. faecal/urine slurry samples, bulk milk samples, and milk filters from dairy herds were negative for e. coli o157.h7. e. coli o157.h7 was also isolated from 10 of 1412 (0.71%) faecal samples from pastured beef cattle in 4 of 25 (16%) herds. the prevalence of e. coli o1 ... | 1994 | 7925659 |
| further oxidation of hydroxycalcidiol by calcidiol 24-hydroxylase. a study with the mature enzyme expressed in escherichia coli. | the coding region of the cdna for rat kidney calcidiol 24-hydroxylase (p450cc24), which is involved in calcium homeostasis in animals, was inserted into an expression vector pkk223-3. the recombinant plasmid was formed in a specific manner without deletion or substitution of any parts of the coding region of the cdna. when the resulting plasmid was introduced into escherichia coli jm109, the recombinant cells produced a protein which was immunoreactive to an antibody against p450cc24. when the c ... | 1994 | 7925346 |
| isolation of escherichia coli synthesized recombinant eukaryotic proteins that contain epsilon-n-acetyllysine. | recombinant porcine (rpst) and bovine somatotropins (rbst) synthesized in escherichia coli contain the amino acid, epsilon-n-acetyllysine. this amino acid was initially discovered in place of the normal lysine144 in a modified reversed-phase hplc (rp-hplc) species of rpst. mass spectrometry and amino acid sequencing of a tryptic peptide isolated from this rp-hplc purified protein were used to identify this altered residue as epsilon-n-acetyllysine. ion-exchange chromatography was utilized to pre ... | 1994 | 7920255 |
| recombinant human erythrocyte cytochrome b5. | the gene encoding the human erythrocyte form of cytochrome b5 (97 residues in length) has been prepared by mutagenesis of an expression vector encoding lipase-solubilized bovine liver microsomal cytochrome b5 (93 residues in length) (funk et al., 1990). efficient expression of this gene in escherichia coli has provided the first opportunity to obtain this protein in quantities sufficient for physical and functional characterization. comparison of the erythrocytic cytochrome with the trypsin-solu ... | 1994 | 7918357 |
| protective antibody titres and antigenic competition in multivalent dichelobacter nodosus fimbrial vaccines using characterised rdna antigens. | the relationship between k-agglutination antibody titres and protection against experimental challenge with dichelobacter nodosus, the effect of increasing the number of d. nodosus fimbrial antigens, and the importance of the nature of additional antigens in multivalent vaccines on antibody response and protection against experimental challenge with d. nodosus were examined in merino sheep. a total of 204 merino sheep were allocated to one of 12 groups, and vaccinated with preparations containin ... | 1994 | 7909183 |
| hemagglutinating properties of enteroaggregative escherichia coli. | many intestinal bacterial pathogens possess hemagglutinating properties, which are indicative of their adhesive properties to the intestinal mucosal surface. to understand the bacteria-mucosa interaction, 41 strains of enteroaggregative escherichia coli (eaggec), a recently described category of diarrheagenic e. coli, isolated mostly from children with diarrhea in bangladesh, india, thailand, central america, and south america were screened for mannose-sensitive hemagglutination and mannose-resi ... | 1994 | 7908675 |
| research note: avidity of chicken yolk antibodies to enterotoxigenic escherichia coli fimbriae. | the yolk antibodies from chickens and the serum and colostrum antibodies from cows were obtained after immunization of these animals with inactivated bacterin or purified k99 fimbriae from enterotoxigenic escherichia coli (etec). the avidity of anti-k99 fimbriae antibodies produced from either chickens or cows was measured by competitive binding assay of elisa. the yolk antibodies competed strongly with the serum and colostrum antibodies from immunized cows and inhibited 40 to 80% of the binding ... | 1993 | 7906035 |
| truncated groel monomer has the ability to promote folding of rhodanese without groes and atp. | similar to chaperonins from other sources, intact chaperonin from escherichia coli (groel) exists as a tetradecamer, and the ability to promote folding of other proteins has been considered to be dependent on this oligomeric structure. however, the peptide fragments of groel of molecular size 34-50 kda, which are produced by limited proteolysis of monomeric groel and are unable to assemble into an oligomer, retain the ability to promote folding of rhodanese even though the yield of productive fo ... | 1993 | 7903258 |
| expression of cytochrome p450 3a5 in escherichia coli: effects of 5' modification, purification, spectral characterization, reconstitution conditions, and catalytic activities. | cytochrome p450 (p450) 3a5 is a human enzyme with 85% amino acid sequence identity to the more predominantly expressed p450 3a4 and has been reported to have overlapping catalytic specificity. the 5'-terminus of a p450 3a5 cdna was modified for optimal expression in escherichia coli using the vector pcw, by aligning the malllavfl n-terminal sequence of recombinant bovine p450 17a (h. j. barnes, m. p. arlotto, and m. r. waterman, (1991) proc. natl. acad. sci. usa 88, 5597-5601) to the 3a5 cdna. t ... | 1995 | 7893152 |
| immunocytochemical localization of heterologously expressed adrenodoxin and its electron acceptor cytochrome p45011b1 in escherichia coli. | adrenal steroid hydroxylase p45011b1 and its electron donor adrenodoxin were localized in the cortex of bovine adrenals by immunogold-silver staining. in order to test recently developed heterologous expression systems for both enzymes to enable structure-function studies, immunocytochemical marker methods were applied. adrenodoxin, the ferredoxin of the adrenal gland, was successfully expressed and for the first time localized in escherichia coli. by use of ultrathin cryosections and the protei ... | 1994 | 7889992 |
| detection of verotoxigenic escherichia coli in bull semen using the polymerase chain reaction. | oligonucleotide primers used in a polymerase chain reaction (pcr) protocol detected the verotoxin 2 (vt2) gene in e. coli present in experimentally contaminated bull semen. the vt2 (shiga-like toxin ii [slt-ii]) primers targeted a 346-bp fragment of the gene coding for the a subunit of the toxin. pcr products, corresponding to the vt2 gene sequence, were amplified from template e. coli nucleic acid extracted from 18-h broth culture and from e. coli in contaminated semen in the undiluted state, d ... | 1994 | 7886935 |
| high incidence of haemorrhagic colitis due to escherichia coli o157 in one scottish town: clinical and epidemiological features. | verotoxin-producing strains of escherichia coli (vtec), in particular serotype o157:h7, are now recognised as the major cause of haemorrhagic colitis and the haemolytic uraemic syndrome (hus) in the u.k. and in north america, and increasingly so in other countries. over a 3-year period (1989-1991), 16 cases of e. coli 0157 infection occurred in one town (peterhead) in north-east grampian. four patients required admission to hospital, of whom three developed hus. the bovine source of vtec infecti ... | 1994 | 7884230 |
| laboratory investigation of a multistate food-borne outbreak of escherichia coli o157:h7 by using pulsed-field gel electrophoresis and phage typing. | two hundred thirty-three isolates of escherichia coli o157:h7 were analyzed by both pulsed-field gel electrophoresis (pfge) and bacteriophage typing. all 26 isolates from persons whose illness was associated with a recent multistate outbreak of e. coli o157:h7 infections linked to the consumption of undercooked hamburgers and all 27 isolates from incriminated lots of hamburger meat had the same phage type and the same pfge pattern. twenty-five of 74 e. coli o157:h7 isolates from washington state ... | 1994 | 7883892 |
| an actin-related protein from dictyostelium discoideum is developmentally regulated and associated with mitochondria. | an actin-related protein (acla) has been identified in the cellular slime mould dictyostelium discoideum. the complete cdna sequence indicates that within the actin superfamily it belongs to the arp3 family of actin-related proteins together with arp66b from drosophila melanogaster, actin2 from bos taurus, act2 from schizosaccharomyces pombe and possibly act2 from caenorhabditis elegans. the acla mrna is regulated during development, showing a maximum between 2 and 4 h after starvation. the prot ... | 1995 | 7883039 |
| enteric pathogens in intensively reared veal calves. | observations were made on development of diarrhea in special-fed calves (n = 460) on 8 commercial facilities during 2 successive 16-week production cycles at weeks 0, 2, 4, 8, 12, and 16. a total of 23% were affected, with peak number of calves with diarrhea observed at week 0. suspected enteropathogens were identified in 86% of these calves, most commonly cryptosporidia, coronavirus, and rotavirus. identified potential zoonotic pathogens included giardia and salmonella spp and verotoxigenic esc ... | 1994 | 7879973 |
| solution structure of the dna binding domain of a nucleoid-associated protein, h-ns, from escherichia coli. | the three-dimensional structure of the c-terminal domain (47 residues) obtained from the hydrolysis of h-ns protein with bovine trypsin was determined by nmr measurements and distance geometry calculations. it is composed of an antiparallel beta-sheet, an alpha-helix and a 3(10)-helix which form a hydrophobic core, stabilizing the whole structure. this domain has been found to bind to dna. possible dna binding sites are discussed on the basis of the solution structure of the c-terminal domain of ... | 1995 | 7875316 |
| [participation of the quinone acceptor in the transition of complex i from an inactive to active state]. | almost complete slow activation of the deactivated purified complex i was observed after the steady-state nadh: cytochrome c reductase reaction turnovers catalyzed by the endogenous (tightly-bound) ubiquinone and contaminant complex iii. the rotenone-sensitive nadh oxidase was reconstituted from bovine heart complex i and escherichia coli quinol-oxidase. the ratio between active and inactive complex i during the steady-state nadh oxidase reaction catalyzed by the reconstituted system was shown t ... | 1994 | 7873673 |
| expression of bovine heart fructose 6-phosphate,2-kinase:fructose 2,6-bisphosphatase and determination of the role of the carboxyl terminus by mutagenesis. | bovine heart fructose 6-p,2-kinase:fructose 2,6-bisphosphatase was expressed in escherichia coli. in order to determine the role of the carboxyl-terminal peptide, 49 and 78 amino acids from the c-terminus were deleted using oligonucleotide-directed mutagenesis. the expressed wild-type and mutant enzymes were purified to homogeneity, and the steady-state kinetics of the mutant enzymes were compared to those of the wild-type enzyme. deletion of 49 residues (del 49) resulted in a 35% decrease in km ... | 1995 | 7873535 |
| use of a recombinant antigen in an indirect elisa for detecting bovine antibody to capripoxvirus. | the gene coding for the capripoxvirus structural protein p32 was cloned, expressed in escherichia coli as a fusion protein with glutathione-s-transferase, and purified on glutathione sepharose. an indirect enzyme linked immunosorbent assay (elisa) using this antigen was developed to screen bovine sera for antibodies to capripoxvirus. sequential serum samples from experimentally infected animals tested by elisa and by virus neutralisation test (vnt) showed that the elisa was more sensitive and de ... | 1994 | 7868646 |
| optimal enzymatic hydrolysis of urinary benzodiazepine conjugates. | conditions for the enzymatic hydrolysis of benzodiazepine conjugates were systematically examined. optimal recovery of the free drugs occurs when 1 ml of urine buffered to ph 4.5 is incubated with 5000 u of helix pomatia beta-glucuronidase at 56 degrees c for 2 h. urine specimens containing conjugates of temazepam, oxazepam, lorazepam, alpha-hydroxyalprazolam, 2-hydroxyethylflurazepam, and n-desalkyl-3-hydroxyflurazepam as targeted benzodiazepines were used. the freed drugs were quantitated by g ... | 1994 | 7861750 |
| enhanced binding of enterotoxigenic escherichia coli k99 to amide derivatives of the receptor ganglioside neugc-gm3. | a natural receptor in pig small intestine [teneberg, s., willemsen, p., de graaf, f. k., & karlsson, k.-a. (1990) febs lett. 263, 10-14] for the enterotoxigenic bacteria escherichia coli k99 is the ganglioside neugc-gm3 (neugc alpha 3gal beta 4glc beta cer) [e.g., h. smit, w. gaastra, j. p. kamerling, j. f. g. vliegenthart, & f. k. de graaf (1984) infect. immun. 46, 578-584]. chemical modifications of the carboxyl group of this ganglioside were performed, giving five different amides, the methyl ... | 1995 | 7849044 |
| [localization of gh gene to bovine chromosome 5q22-26 by in situ hybridization]. | the bgh entire gene (3.0kb), as a probe which was cloned into recombinant plasmid pbgh was used for in situ hybridization. the e. coli rr1, as a receptor, was transformed by recombinant plasmid pbgh. recombinant plasmid dnas were amplified, extracted and purified, the bgh fragments were collected and labelled by nicktranslation. the metaphase and early-metaphase chromosome spreads were prepared from tdr-brdu-synchronized peripheral blood lymphocytes in beijing black-white dairy cattle. after in ... | 1994 | 7848663 |
| sequence and expression of the ns2 protein gene of human coronavirus oc43. | the complete nucleotide sequence of the ns2 gene of human coronavirus oc43 (hcv-oc43) was determined. sequence analysis revealed an open reading frame that could encode a protein of 278 amino acids, with an estimated molecular mass of 32.2 kda. six potential phosphorylation sites are present but no sites of n-glycosylation were found. the amino acid sequence of the hcv-oc43 ns2 protein shows 92% identity with that of the mebus strain of bovine coronavirus (bcv). however, a stretch of nine consec ... | 1995 | 7844564 |
| energy-transducing nicotinamide nucleotide transhydrogenase: nucleotide sequences of the genes and predicted amino acid sequences of the subunits of the enzyme from rhodospirillum rubrum. | based on the amino acid sequence of the n-terminus of the soluble subunit of the rhodospirillum rubrum nicotinamide nucleotide transhydrogenase, two oligonucleotide primers were synthesized and used to amplify the corresponding dna segment (110 base pairs) by the polymerase chain reaction. using this pcr product as a probe, one clone with the insert of 6.4 kbp was isolated from a genomic library of r. rubrum and sequenced. this sequence contained three open reading frames, constituting the genes ... | 1994 | 7844118 |
| effects of housing and colostrum feeding on the prevalence of selected infectious organisms in feces of jersey calves. | neonatal jersey calves (n = 96) were used to evaluate effects of housing (individual hutches or wooden pens in a barn) and colostrum feeding (calves were separated from the dam and fed 2 l of colostrum in nipple-bottles or allowed to nurse the dam for 3 d) on the prevalence of selected organisms in feces. prevalence of cryptosporidium and eimeria were reduced, and prevalence of rotavirus tended to be reduced, when calves were housed in hutches. prevalence of coronavirus was unaffected by treatme ... | 1994 | 7836601 |
| a group of novel glutathione s-transferase isozymes showing high activity towards 4-hydroxy-2-nonenal are present in bovine ocular tissues. | recently, a mouse glutathione s-transferase (gst) isozyme, mgsta4-4, which belongs to a distinct group of gsts has been characterized in our laboratory. during the present studies, western blot analyses of bovine ocular tissues using the antibodies raised against the recombinant mgsta4-4 obtained by expression in escherichia coli revealed that the orthologs of mgsta4-4 were present in cornea, retina, iris-ciliary body and sclera, but absent in lens. these novel gst isozymes of bovine ocular tiss ... | 1994 | 7835404 |
| brucella abortus serology in cattle naturally infected with escherichia coli o157.h7. | 1994 | 7831743 | |
| biological activities and secondary structures of variant forms of human salivary cystatin sn produced in escherichia coli. | using an escherichia coli expression system, pgex-2t, that expresses foreign sequences as fusion proteins with a glutathione s-transferase (gst) carrier, we have produced several recombinant human salivary cystatin sn (recsnsn) variants. these include a n-terminal-truncated form (aa 17-121), a c-terminal-truncated form (aa 1-102) and two deletion mutants (delta 12-16 and delta 56-60). a large amount of the insoluble fusion protein (approx. 15 mg/l) was produced in each case. these were solubiliz ... | 1994 | 7828895 |
| the effect of low temperatures on enzyme activity. | the stability of two enzymes from extreme thermophiles (glutamate dehydrogenase from thermococcales strain an1 and beta-glucosidase from caldocellum saccharolyticum expressed in escherichia coli) has been exploited to allow measurement of activity over a 175 degrees c temperature range, from +90 degrees c to -85 degrees c for the glutamate dehydrogenase and from +90 degrees c to -70 degrees c for the beta-glucosidase. the arrhenius plots of these enzymes, and those for two mesophilic enzymes (gl ... | 1995 | 7826325 |
| characterization of a mammalian homolog of the escherichia coli muty mismatch repair protein. | a protein homologous to the escherichia coli muty protein, referred to as myh, has been identified in nuclear extracts of calf thymus and human hela cells. western blot (immunoblot) analysis using polyclonal antibodies to the e. coli muty protein detected a protein of 65 kda in both extracts. partial purification of myh from calf thymus cells revealed a 65-kda protein as well as a functional but apparently degraded form of 36 kda, as determined by glycerol gradient centrifugation and immunoblott ... | 1995 | 7823963 |
| induction of inflammatory cytokines in bovine alveolar macrophages following stimulation with pasteurella haemolytica lipopolysaccharide. | bovine tumor necrosis factor alpha (tnf-alpha) and interleukin-1 beta (il-1 beta) cdnas were generated by reverse transcription and then by pcr amplification from lipopolysaccharide (lps)-stimulated alveolar macrophage rna. the amplified cdnas were cloned into ppow and expressed in escherichia coli dh5 alpha. the expressed proteins were confirmed as tnf-alpha and il-1 beta by western blot (immunoblot) analysis and bioassays. we then used the cloned genes as probes in northern (rna) blots and inv ... | 1995 | 7822000 |
| coliform counts and escherichia coli in raw commercial milk from dairy farmers in kiambu district, kenya. | the rate of contamination with coliforms and incidence of escherichia coli (e. coli) in raw milk supplied by farmers to dairy cooperative societies for marketing was investigated. about forty two (42.2%) percent of the milk samples from farmers cans and 10.3% of samples from cooperative cans were found to be free of coliforms, while 89.5% of the samples from farmers cans and 50% samples from cooperative cans could be considered to be of good quality with no more than 50,000 coliforms/ml of milk. ... | 1994 | 7821241 |
| application of monoclonal antibody-based sandwich elisas to detect verotoxins in cattle faeces. | b1p4wich elisas, which use monoclonal antibodies for both the capture and the completion stages, were developed for the detection of verotoxins i and ii (non-variant). the elisas were used to investigate the incidence of verotoxins in 304 samples of bovine faeces collected from cases of enteritis, and 113 samples collected from normal animals. for most samples, the tests were carried out directly on faeces, and on colony sweeps which were taken from cultures grown from the faeces. these colonies ... | 1994 | 7817011 |
| inositol hexakisphosphate binds to clathrin assembly protein 3 (ap-3/ap180) and inhibits clathrin cage assembly in vitro. | we have isolated an inositol hexakisphosphate binding protein from rat brain by affinity elution chromatography from mono s cation exchange resin using 0.1 mm inositol hexakisphosphate (insp6). the amino acid sequences of six tryptic peptides from the protein were identical to the sequences predicted from the cdna encoding a previously isolated protein designated as ap-3 or ap180. this protein is localized in nerve endings and promotes assembly of clathrin into coated vesicles. the isolated prot ... | 1995 | 7814377 |
| expression and folding of recombinant bovine prethrombin-2 and its activation to thrombin. | bovine prethrombin-2 has been produced in escherichia coli using a t7 expression system. the expressed prethrombin-2 formed intracellular inclusion bodies which were solubilized by reversible sulfonation of the cysteines in the presence of 7 m guanidine hydrochloride. sulfonated prethrombin-2 was refolded in the presence of 4 m guanidine hydrochloride, using oxidized and reduced glutathione as the redox couple. the folded protein was purified by heparin affinity chromatography and activated to t ... | 1995 | 7814368 |
| [detection of verotoxin-producing e. coli in field isolates from domestic and agricultural animals in sachsen-anhalt]. | a report is given on the detection of verotoxin-producing e. coli (vtec) strains from field isolates of healthy or ill cattle (n = 141), pigs (n = 306), sheep (n = 15), cats (n = 29) and dogs (n = 25) in the region of the new federal land sachsen-anhalt. 5% of the strains isolated from cattle, 32% from pigs, 20% from sheep, 4% from dogs and 0% from cats have shown vtec. the e. coli-strains were checked for the presence of other factors of virulence, too. a good correlation (82%) was found betwee ... | 1994 | 7802620 |
| direct association of occludin with zo-1 and its possible involvement in the localization of occludin at tight junctions. | occludin is an integral membrane protein localizing at tight junctions (tj) with four transmembrane domains and a long cooh-terminal cytoplasmic domain (domain e) consisting of 255 amino acids. immunofluorescence and laser scan microscopy revealed that chick full-length occludin introduced into human and bovine epithelial cells was correctly delivered to and incorporated into preexisting tj. further transfection studies with various deletion mutants showed that the domain e, especially its cooh- ... | 1994 | 7798316 |
| determination of peptide regions exposed at the surface of the bacterial ribosome with antibodies against synthetic peptides. | we synthesized six peptides corresponding to regions that are predicted to be surface-exposed of the following ribosomal proteins: protein l2, positions (d263-k272); protein l5, positions (i136-g150); protein l25, positions (q75-d90); protein s3, positions (q222-k232) derived from escherichia coli; and protein l2, positions (k257-k275), and protein s3, positions (r130-t150) from bacillus stearothermophilus. these peptides were employed to raise ribosomal protein-cross-reactive antibodies. the an ... | 1995 | 7794529 |
| characterization of a recombinant pneumolysin and its use as a protein carrier for pneumococcal type 18c conjugate vaccines. | pneumolysin from streptococcus pneumoniae was expressed in escherichia coli as a glutathione s-transferase fusion protein and purified by affinity and hydroxylapatite chromatography. the purified recombinant pneumolysin (rpl), with a molecular mass of 53 kda, had a specific activity of 3 x 10(5) hemolytic units per mg of protein on rabbit erythrocytes and reacted identically in immunodiffusion with the antisera against native pneumolysin. the rpl was used as a protein carrier to prepare conjugat ... | 1995 | 7790088 |
| hemolytic uremic syndrome in children. | the majority of postdiarrheal hemolytic uremic syndrome is caused by escherichia coli o157:h7. the organisms are carried in the intestines of cattle; partially cooked contaminated hamburger is the single most common vector. the e. coli produce a potent cytotoxin that gains access to the circulation, is taken up by glycolipid receptors on glomerular endothelial cells, is internalized, and causes cell death. associated phenomena include the activation of platelets, leukocytes, and the coagulation ... | 1995 | 7787931 |
| ceftiofur distribution in serum and milk from clinically normal cows and cows with experimental escherichia coli-induced mastitis. | eight holstein cows, 4 inoculated intracisternally in 1 quarter of the mammary gland with escherichia coli and 4 noninfected controls, were administered ceftiofur sodium (3 mg/kg of body weight, iv, q 12 hours) for 24 hours, beginning at 14 hours after inoculation of infected cows. all challenge-exposed cows became infected, with mean +/- sem peak log10 bacterial concentration in milk of 5.03 +/- 0.69 colony-forming units/ml. the infection resulted in systemic signs (mean peak rectal temperature ... | 1995 | 7785826 |
| myxococcus xanthus, a gram-negative bacterium, contains a transmembrane protein serine/threonine kinase that blocks the secretion of beta-lactamase by phosphorylation. | a gene, pkn2, encoding a myxococcus xanthus protein with significant similarities to eukaryotic protein serine/threonine kinases, was cloned using the polymerase chain reaction. the open reading frame for the protein, beginning with a gug initiation codon, consists of 830 amino acids. the amino-terminal 279 residues show 37% identity to catalytic domain of pkn1, another protein serine/threonine kinase expressed during the development at the onset of sporulation. the catalytic domain of pkn2 cont ... | 1995 | 7774814 |
| streptococcal protein mag--a protein with broad albumin binding specificity. | protein mag is a cell surface protein from streptococcus dysgalactiae which binds alpha 2-macroglobulin (alpha 2m), serum albumin and immunoglobulin g (igg). in this work protein mag was expressed in escherichia coli, purified and analysed for its albumin-binding specificity. the binding of protein mag to serum albumins of different species origin was studied in a dot-blot assay and compared with the binding of streptococcal protein g, so far the best studied bacterial albumin receptor. the albu ... | 1995 | 7766685 |
| inhibitory effects of milk gangliosides on the adhesion of escherichia coli to human intestinal carcinoma cells. | the effects of milk gangliosides and their derivatives on the adhesion of enterotoxigenic and enteropathogenic escherichia coli to caco-2 cells, a human intestinal carcinoma cell line, were investigated. human milk gangliosides inhibited the adhesion of enterotoxigenic e. coli to caco-2 cells in the same proportion, regardless of the lactational stage, but bovine milk gangliosides were less effective. the most effective inhibitor was monosialoganglioside 1 (gm1); the adhesion rate of enterotoxig ... | 1995 | 7765978 |
| a carbohydrate biosensor surface for the detection of uropathogenic bacteria. | we have developed a new surface for use in biosensors that is based on a gold plate covered with a specific carbohydrate receptor structure. the carbohydrate, gal alpha 1-4gal, was bound covalently via a thioalkylcarboxy-spacer, or adsorbed as a neoglycoprotein, to a two-dimensional gold surface. both types of surfaces showed high specificity in the binding of the uropathogenic bacteria p-fimbriated escherichia coli compared to the binding of non-infectious bacteria. the signal to noise ratio is ... | 1994 | 7765569 |
| high level secretion of calf chymosin using a glucoamylase-prochymosin fusion gene in aspergillus oryzae. | a recombinant chymosin was secreted at high levels using fusion genes with a. oryzae glucoamylase gene (glaa) and a wheat bran solid-state culture system. two portions of the a. oryzae glucoamylase, one with almost the entire glucoamylase (ga1-603) lacking 9 amino acids at the carboxyl terminal, and the other (ga1-511) lacking the starch binding-domain, were fused in frame with prochymosin cdna. western blot analysis indicated that the mature chymosin was released from the secreted fusion protei ... | 1994 | 7764977 |