Publications
| Title | Abstract | Year Filter | PMID(sorted descending) Filter |
|---|
| immunocharacterization of vitis vinifera l. ferredoxin-dependent glutamate synthase, and its spatial and temporal changes during leaf development. | the grapevine (vitis vinifera l.) partial fragment of cdna clone pgogat1 [loulakakis and roubelakis-angelakis (1997) physiol plant 101:220-228], encoding the ferredoxin-dependent glutamate synthase (fd-gogat; ec 1.4.7.1), was overexpressed in escherichia coli cells. a hybrid between the fd-gogat fragment and maltose-binding protein was purified and used to raise a polyclonal antibody in a rabbit. the prepared antibody appeared to be specific towards fd-gogat; it recognized a protein band of appr ... | 2002 | 12172846 |
| development of a highly sensitive in vitro assay method for biological activity of endotoxin contamination in biological products. | the pyrogen test in rabbits has been replaced by the bacterial endotoxin test. the endotoxin test, however, showed a considerable discrepancy with pyrogenicity and was, therefore, assumed to have an efficacy limitation in directly predicting harmful biological effects of endotoxin. we developed a sensitive in vitro assay method by making use of tumour necrosis factor alpha (tnf-alpha) induction in raw264.7 cells, which showed a fine correlation with pyrogenicity in rabbits. raw264.7 cells mainta ... | 2002 | 12127309 |
| regulation of squalene synthase, a key enzyme of sterol biosynthesis, in tobacco. | squalene synthase (ss) represents a putative branch point in the isoprenoid biosynthetic pathway capable of diverting carbon flow specifically to the biosynthesis of sterols and, hence, is considered a potential regulatory point for sterol metabolism. for example, when plant cells grown in suspension culture are challenged with fungal elicitors, suppression of sterol biosynthesis has been correlated with a reduction in ss enzyme activity. the current study sought to correlate changes in ss enzym ... | 2002 | 12114564 |
| molecular cloning and characterization of a new japanese cedar pollen allergen homologous to plant isoflavone reductase family. | japanese cedar (cryptomeria japonica) pollen is a major cause of seasonal pollinosis, and more than 10% of japanese people suffer from this allergic disorder. however, only two major pollen allergens, cry j 1 and cry j 2, have been identified and exclusively characterized. | 2002 | 12100055 |
| cysteine biosynthetic enzymes are the pieces of a metabolic energy pump. | understanding the mechanisms of free energy transfer in metabolism is fundamental to understanding how the chemical forces that sustain the molecular organization of the cell are distributed. recent studies of molecular motors (1-3) and atp-driven proton transport (4-6) describe how chemical potential is transferred at the molecular level. these systems catalyze energy transfer through structural change and appear to be dedicated exclusively to their coupling tasks (7, 8). here we report the dis ... | 2002 | 12081500 |
| expression and purification of the recombinant enteropathogenic escherichia coli vaccine candidates bfpa and espb. | bfpa, the structural repeating protein subunit a of the bundle-forming pilus and espb, a type-iii-secreted pore-forming protein of enteropathogenic escherichia coli (epec), both virulence factors central for epec pathogenesis, were overexpressed in e. coli dh5alpha and m15 laboratory strains, respectively, using the pqe-30 cloning expression system, as chimeric fusions to a nh(2)-terminal histidine hexapeptide (his(6)-tag) sequence. after isopropyl beta-d-thiogalactoside induction, the expressio ... | 2002 | 12071694 |
| expression, purification, and evidence for the interaction of the two nucleotide-binding folds of the sulphonylurea receptor. | the atp-sensitive potassium channel is made up of four pore forming kir6.2 subunits, surrounded by four regulatory sulphonylurea receptor (sur) subunits. the latter subunit contains two nucleotide-binding folds (nbfs) that confer the ability on the channel to sense changes in the metabolic status ([atp]/[adp]) of the cell and couple the changes to the membrane potential of the cell. in an attempt to better understand the mechanisms by which nbfs influence the activity of the channel, we have exp ... | 2002 | 12054762 |
| expression of cd14 protein and its gene in liver sinusoidal endothelial cells during endotoxemia. | to observe expression of cd14 protein and cd14 gene in rat liver sinusoidal endothelial cells (lsecs) during endotoxemia, and the role of cd14 protein in the activation of lipopolysaccharide (lps)-induced lsecs. | 2002 | 12046090 |
| [surveillance of escherichia coli o157:h7 among animals in jiangsu province in 1999]. | to understand the escherichia coli o157:h7 carrier rate of host animals and the toxic gene of the strains in different areas in jiangsu province. | 2002 | 12015089 |
| immunolocalization of 1- o-sinapoylglucose:malate sinapoyltransferase in arabidopsis thaliana. | the serine carboxypeptidase-like protein 1- o-sinapoylglucose:malate sinapoyltransferase (smt) catalyzes the transfer of the sinapoyl moiety of 1- o-sinapoylglucose to malate in the formation of sinapoylmalate in some members of the brassicaceae. rabbit polyclonal monospecific antibodies were raised against the recombinant smt produced in escherichia coli from the corresponding arabidopsis thaliana (l.) heynh. cdna. immunoblot analysis of protein from different arabidopsis tissues showed that th ... | 2002 | 12012238 |
| purification, characterization, and immunogenicity of the refolded ectodomain of the plasmodium falciparum apical membrane antigen 1 expressed in escherichia coli. | the apical membrane antigen 1 (ama1) has emerged as a promising vaccine candidate against malaria. advanced evaluation of its protective efficacy in humans requires the production of highly purified and correctly folded protein. we describe here a process for the expression, fermentation, refolding, and purification of the recombinant ectodomain of ama1 (amino acids 83(gly) to 531(glu)) of plasmodium falciparum (3d7) produced in escherichia coli. a synthetic gene containing an e. coli codon bias ... | 2002 | 12011004 |
| polyclonal antibodies to glutathione s-transferase--verotoxin subunit a fusion proteins neutralize verotoxins. | the a1 subunits of verotoxin-1 (vt1) and vt2 genes were cloned into pgex-4t-2 for the expression of glutathione s-transferase (gst) fusion proteins. the n-terminal and the transmembrane regions of the a1 subunits were excluded from the constructs in order to increase the product yields. polyclonal anti-vt1a1 and anti-vt2a1 antibodies were produced by immunizing rabbits with gst-vt1a1 and gst-vt2a1 fusion proteins, respectively. the antibodies were tested for their ability to neutralize active to ... | 2002 | 11986278 |
| mutation of conserved active-site threonine residues in creatine kinase affects autophosphorylation and enzyme kinetics. | muscle-type creatine kinase (mm-ck) is a member of an isoenzyme family with key functions in cellular energetics. it has become a matter of debate whether the enzyme is autophosphorylated, as reported earlier [hemmer, furter-graves, frank, wallimann and furter (1995) biochim. biophys. acta 1251, 81-90], or exclusively nucleotidylated. in the present paper, we demonstrate unambiguously that ck is indeed autophosphorylated. however, this autophosphorylation is not solely responsible for the observ ... | 2002 | 11964180 |
| novel anticoagulant activity of polybrene: inhibition of monocytic tissue factor hypercoagulation following bacterial endotoxin induction. | the enhanced extrinsic coagulation in response to inflammation could contribute to disseminated intravascular coagulation, often manifesting cardiovascular complications. the complex mechanism remains unclear. nor is the effective anticoagulation well established. the search for arresting hypercoagulation is of antithrombotic relevance. the ability of polybrene (pb) to inhibit tissue factor (tf)-initiated extrinsic blood coagulation was demonstrated at the protein and cellular levels as well as ... | 2002 | 11914654 |
| subcellular localization of chlorosome proteins in chlorobium tepidum and characterization of three new chlorosome proteins: csmf, csmh, and csmx. | chlorosomes are unique light-harvesting structures found in two families of photosynthetic bacteria. in this study, three chlorosome proteins (csmf, csmh, and csmx) of the green sulfur bacterium chlorobium tepidum were characterized by cloning and sequencing the genes which encode them, by overproducing the respective proteins in escherichia coli, and by raising polyclonal antisera to the purified proteins. three other proteins (atpf, ct1970, and ct2144) which were identified in chlorosome fract ... | 2002 | 11914082 |
| effects of coligand variation on the in vivo characteristics of tc-99m-labeled interleukin-8 in detection of infection. | in our previous studies, interleukin-8 (il-8) was labeled with (99m)tc using hydrazinonicotinamide (hynic) as bifunctional coupling agent and tricine as coligand. this preparation showed excellent characteristics for imaging of infection in a rabbit model of soft-tissue infection. in the present study, the propylaldehyde hydrazone formulation of hynic was introduced to stabilize hynic-il-8. (99m)tc-hynic-il-8 was prepared using 5 different coligand formulations. the effect of these coligand form ... | 2002 | 11906276 |
| cross-protection against challenge by intravenous escherichia coli verocytotoxin 1 (vt1) in rabbits immunized with vt2 toxoid. | rabbits challenged intravenously with escherichia coli verocytotoxin (vt1, shiga toxin 1, stx1) die after developing diarrhea and paralysis, and this outcome can be prevented by pre-immunization with vt1 toxoid. in nonimmune rabbits, intravenously administered 125i-vt1 binds to the central nervous system and gastrointestinal tract, whereas in immunized animals, these organs are spared and the toxin localizes in the liver and spleen. in rabbits immunized with either vt1 or vt2 toxoids, both the h ... | 2002 | 11888169 |
| a monoclonal antibody to tryptophan hydroxylase: applications and identification of the epitope. | recombinant rabbit tryptophan hydroxylase (tph) was expressed in escherichia coli and purified from inclusion bodies by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page). a mouse monoclonal antibody and rabbit and sheep polyclonal antibodies were generated. in immunohistochemical studies of formaldehyde-fixed primate brain, the monoclonal strongly labeled not only cell bodies in the raphe nuclei but also fibers in the cerebral cortex. truncation mutants and peptide ... | 2002 | 11856572 |
| contribution of fibronectin-binding protein to pathogenesis of streptococcus suis serotype 2. | in the present study we investigated the role of the fibronectin (fn)- and fibrinogen (fgn)-binding protein (fbps) in the pathogenesis of streptococcus suis serotype 2 in piglets. the complete gene encoding fbps from s. suis serotype 2 was cloned in escherichia coli and sequenced. the occurrence of the gene in various serotypes was analyzed by hybridization studies. the fbps protein was expressed in e. coli and purified, and binding to human fn and fgn was demonstrated. the induction of antibodi ... | 2002 | 11854216 |
| two mutations in troponin i that cause hypertrophic cardiomyopathy have contrasting effects on cardiac muscle contractility. | we investigated the effects of two mutations in human cardiac troponin i, arg(145)-->gly and gly(203)-->ser, that are reported to cause familial hypertrophic cardiomyopathy. mutant and wild-type troponin i, overexpressed in escherichia coli, were used to reconstitute troponin complexes in vanadate-treated guinea pig cardiac trabeculae skinned fibres, and thin filaments were reconstituted with human cardiac troponin and tropomyosin along with rabbit skeletal muscle actin for in vitro motility and ... | 2002 | 11853553 |
| stimulation of mucosal immune response following oral administration of enterotoxigenic escherichia coli fimbriae (cfa/i) entrapped in liposomes in conjunction with inactivated whole-cell vibrio cholerae vaccine. | in this study, we have searched for an effective mucosal vaccine. an oral enterotoxigenic e. coli vaccine containing colonization factor antigen (cfa/i) associated with inactivated whole-cell v. cholerae vaccine (wcv) has been tested for safety and immunogenicity in animals. five groups of animals were used. the results showed the following: (a) vaccine containing cfa/i antigen entrapped in liposomes and associated with wcv (batch c) had increased titers of specific antibodies to cfa/i antigen i ... | 2001 | 11850896 |
| effects of lipopolysaccharide on low- and high-density cultured rabbit vascular smooth muscle cells: differential modulation of nitric oxide release, erk1/erk2 map kinase activity, protein tyrosine phosphatase activity, and dna synthesis. | previous studies have shown that exogenously generated nitric oxide (no) inhibits smooth muscle cell proliferation. in the present study, we stimulated rabbit vascular smooth muscle cells (rvsmc) with e. coli lipopolysaccharide (lps), a known inducer of no synthase transcription, and established a connection between endogenous no, phosphorylation/dephosphorylation-mediated signaling pathways, and dna synthesis. non-confluent rvsmc were cultured with 0, 5, 10, or 100 ng/ml of the endotoxin. no re ... | 2002 | 11847521 |
| immune response to escherichia coli antigens entrapped in liposomes. i. immunopathological studies. | in this study, we have searched for an effective mucosal delivery system for a purified e. coli antigen which elicits anticolonization and anti-toxic immunity. e. coli colonization factor antigen (cfa/i) and heat-labile enterotoxin (lt) were encapsulated in liposomes. to determine the efficacies of soluble and liposome-encapsulated e. coli antigens young rabbits were mucosally treated with three oral doses of e. coli antigens given 7 days apart. ten days after the last booster, rabbits were oral ... | 1999 | 11845460 |
| sequencing, expression, and characterization of cdna expressed flavin-containing monooxygenase 2 from mouse. | the cdna clone of mouse flavin-containing monooxygenase 2 (fmo2) was obtained as an expressed sequence tag (est) isolated from a female mouse kidney cdna library from the i.m.a.g.e. consortium (i.m.a.g.e. cloneid 1432164). complete sequencing of the est derived a nucleotide sequence for mouse fmo2, which contains 112 bases of 5' flanking region, 1607 bases of coding region, and 309 bases of 3' flanking region. this fmo2 sequence encodes a protein of 535 amino acids including two putative pyropho ... | 2001 | 11835629 |
| identification and characterization of assembly proteins of cs5 pili from enterotoxigenic escherichia coli. | this study investigated the role of three genes comprising part of the operon which encodes cs5 pili from enterotoxigenic escherichia coli. in-frame gene deletions were constructed, and the effects on biogenesis of the pili were examined. a deletion in csfb abolished csfa major subunit accumulation in the periplasm, which could be restored by trans-complementation with a complete copy of the csfb gene. localization studies using an antibody against csfb showed that this protein was periplasmical ... | 2002 | 11807067 |
| [a study on the pathogenesis of streptococcus mitis exotoxin]. | to study the isolation, purification and pathogenesis of streptococcus mitis pyrogenic exotoxin causing toxic shock syndrome. | 2001 | 11798555 |
| characterization of two genes encoding ferritin-like protein in actinobacillus actinomycetemcomitans. | two genes encoding ferritin-like protein, designated afna and afnb, were identified in the upstream region of actx on the actinobacillus actinomycetemcomitans chromosomal dna. the actx has been reported to be a regulatory gene homologous to the escherichia coli fnr, which controls the growth and virulence of a. actinomycetemcomitans under anaerobic conditions. the afnb located 340 bp-upstream from the actx, and the afna located just 15 bp-upstream from afnb. the afna and afnb encoded 161 and 165 ... | 2001 | 11762755 |
| use of fusion protein constructs to generate potent immunotherapy and protection against scorpion toxins. | we report the use of recombinant scorpion toxins in the form of fusion proteins as antigens for immunisation in rabbits and mice: the aim was to produce in these animal models protective antisera against the most lethal alpha-type toxins in the venom from the north african scorpion androctonus australis. the cdnas encoding aah i, aah ii and aah iii (the three major alpha-type toxins acting on voltage-sensitive sodium channels) were fused to the sequence encoding the maltose binding protein (mbp) ... | 2001 | 11738760 |
| purification of the autotransporter protein hbp of escherichia coli. | the enzyme hbp (hemoglobin protease) of the pathogenic escherichia coli strain eb1 has been purified to homogeneity by gel filtration chromatography. the purified protein is capable of binding heme and shows hemoglobin protease activity. our method of purification is applicable not only to hbp but also to other autotransporter proteins and will contribute to a better understanding of the function-structure relationship of this family of proteins. | 2001 | 11728729 |
| antibiotic resistance in bacteria from magpies (pica pica) and rabbits (oryctolagus cuniculus) from west wales. | the prevalence of antibiotic-resistant bacteria in wild animal and bird populations is largely unknown, with little consistency among the few published reports. we therefore examined intestinal bacteria from magpies (pica pica) and rabbits (oryctolagus cuniculus) collected in rural west wales. escherichia coli isolates resistant to multiple antibiotics were grown from eight of 20 magpies trapped in spring, 1999 and one of 17 in spring, 2000; the most prevalent resistance trait among these isolat ... | 2001 | 11722546 |
| purification of human recombinant gata-1 from bacteria: implication for protein-protein interaction studies. | gata-1 is a key regulator of terminal erythroid differentiation in mammals and birds. the structural and biochemical studies of human gata-1 (hgata-1) are limited by the difficulty of its purification in a sufficient amount. here we describe the procedure for obtaining pure bacterial recombinant hgata-1 in an active functional state. we demonstrate that this protein may be successfully used for preparing an affinity column, producing gata-1-specific rabbit polyclonal antibodies, and studying dna ... | 2001 | 11722179 |
| overexpression, purification, and characterization of glutaminase-interacting protein, a pdz-domain protein from human brain. | a human brain cdna clone coding for a novel pdz-domain protein of 124 amino acids has been previously isolated in our laboratory. the protein was termed gip (glutaminase-interacting protein) because it interacts with the c-terminal region of the human brain glutaminase l. here we report the heterologous expression of gip as a histidine-tagged fusion protein in escherichia coli cells. the induction conditions (temperature and isopropyl beta-d-thiogalactopyranoside concentrations) were optimized i ... | 2001 | 11722177 |
| wild rabbits--a novel vector for verocytotoxigenic escherichia coli o157. | 2001 | 11720214 | |
| molecular characterization of a troponin i-like protein from the hard tick haemaphysalis longicornis. | a cdna expression library prepared from mrna of haemaphysalis longicornis (h. longicornis) was screened with a h. longicornis-infested rabbit serum. a cdna encoding 27/30kda proteins was cloned and designated p27/30 gene. the predicted amino acid sequence of the p27/30 gene shows a rather high homology (58% amino acid identities and 11% amino acid similarity) with drosophila melanogaster troponin i clone e2. h. longicornis p27/30 possesses amino acid sequence of actin-binding domains of troponin ... | 2001 | 11719070 |
| dynamic changes in neutrophil defensins during endotoxemia. | bacterial endotoxin or lipopolysaccharide (lps) is an important causative agent of sepsis. this study determines the expression of defensins np-2 and np-5 and the function of polymorphonuclear leukocytes (pmn) in rabbits treated with lps. pmn functional activity was assessed by measuring cd18 expression and h(2)o(2) production and by examining the lungs. np-2 and, to a minor extent, np-5 of circulating pmn increase during endotoxemia. this early increase is concomitant with neutrophilia and elev ... | 2001 | 11705961 |
| a truncation of 2b subfamily cytochromes p450 yields increased expression levels, increased solubility, and decreased aggregation while retaining function. | the hydrophobic membrane-spanning domain in four cytochromes p450 2b was removed (delta3-21) and several positive charges were substituted at the n-terminus to increase expression and solubility. histidine residues were appended to the c-terminus to simplify purification. the truncated proteins were highly expressed in escherichia coli, could be released from the membrane using high salt conditions, and were purified from this fraction to specific contents up to 19 nmol p450/mg protein using a s ... | 2001 | 11673866 |
| [pathogenic activity modulation of escherichia coli tl+ toxin with an isolated protein of giardia intestinalis and a synthetic peptide]. | it is know that a protein from giardia intestinalis works as a substrate for v. cholerae and escherichia coli. the toxic activity of both activates protein g form intestinal mucosa with a pathogenic activity results. in the present study, the pathogenic activity of subunit a of vibrio cholerae toxin (adp-ribosyltranferase) using isolated fragments from: giardia intestinalis and a synthetic peptide were used as modulators in vivo. | 2001 | 11599482 |
| cloning and expression of a single-chain catalytic antibody that acts as a glutathione peroxidase mimic with high catalytic efficiency. | glutathione peroxidase (gpx) has a powerful role in scavenging reactive oxygen species. in previous papers we have developed a new strategy for generating abzymes: the monoclonal antibody with a substrate-binding site is first prepared, then a catalytic group is incorporated into the monoclonal antibody's binding site by using chemical mutation [luo, zhu, ding, gao, sun, liu, yang and shen (1994) biochem. biophys. res. commun. 198, 1240-1247; ding, liu, zhu, luo, zhao and ni (1998) biochem. j. 3 ... | 2001 | 11583583 |
| transfer of sulfur from iscs to iscu during fe/s cluster assembly. | the cysteine desulfurase enzymes nifs and iscs provide sulfur for the biosynthesis of fe/s proteins. nifu and iscu have been proposed to serve as template or scaffold proteins in the initial fe/s cluster assembly events, but the mechanism of sulfur transfer from nifs or iscs to nifu or iscu has not been elucidated. we have employed [(35)s]cysteine radiotracer studies to monitor sulfur transfer between iscs and iscu from escherichia coli and have used direct binding measurements to investigate in ... | 2001 | 11577100 |
| extremely low doses of tissue factor pathway inhibitor decrease mortality in a rabbit model of septic shock. | we sought to determine the lowest dose of recombinant human tissue factor pathway inhibitor (tfpi) that can provide protection from lethality in a rabbit model of septic shock. | 2001 | 11511939 |
| recombinant listeria strains producing the nontoxic l-chain of botulinum neurotoxin a in a soluble form. | fragments of clostridium botulinum neurotoxin a (bont/a) gene (bota) were expressed in listeria monocytogenes atcc10527 to produce the l-chain of the toxin in a soluble form. a shuttle vector pat19 (emr) was used to make plasmid pat-rl containing a bota gene fragment placed under c. botulinum ntnh-gene promoter control. the plasmid also contained a c. botulinum botr/a gene, a positive transcriptional regulator of bota. the cytoplasmic fraction of the l. monocytogenes (pat-rl) strain was found to ... | 2001 | 11501674 |
| cloning, sequencing, and expression of the leukotoxin gene from fusobacterium necrophorum. | fusobacterium necrophorum is a gram-negative, rod-shaped, anaerobic bacterium that is a primary or secondary etiological agent in a variety of necrotic purulent infections in animals and humans. included are diseases of cattle such as liver abscesses and foot rot, which have economically important consequences for the cattle industry. the major virulence factor of this bacterium is leukotoxin, a secreted protein of high molecular weight active against leukocytes from ruminants. the screening of ... | 2001 | 11500416 |
| characterization of a stress-induced alternate sigma factor, rpos, of coxiella burnetii and its expression during the development cycle. | coxiella burnetii is an obligate intracellular bacterium that resides in an acidified phagolysosome and has a remarkable ability to persist in the extracellular environment. c. burnetii has evolved a developmental cycle that includes at least two morphologic forms, designated large cell variants (lcv) and small cell variants (scv). based on differential protein expression, distinct ultrastructures, and different metabolic activities, we speculated that lcv and scv are similar to typical logarith ... | 2001 | 11447163 |
| production of bartonella genus-specific monoclonal antibodies. | monoclonal antibodies (mabs) which react with heat-resistant proteins with molecular masses of 32 to 33 kda of 14 different bartonella species were produced. these antibodies did not react with antigens of 26 diverse bacterial strains by microimmunofluorescence assay except mab b3d4, which reacted with chlamydia psittaci and chlamydia trachomatis at low titers. the identification of a common bartonella antigenic protein will make it possible to later produce a diagnostic antigen by cloning and e ... | 2001 | 11427441 |
| molecular characterization of a peroxiredoxin from the hard tick haemaphysalis longicornis. | antioxidant enzymes in eukaryotes play an important role in protection against the oxygen radicals generated during aerobic metabolism. here we report the cloning and characterization of a cdna encoding the antioxidant enzyme peroxiredoxin from the hard tick haemaphysalis longicornis (hlprx). hlprx is 939 bp long and contains a 101 bp non-translated sequence at the 5' end and a polyadenylation singnal followed by a poly(a) tail at the 3' end. hlprx encodes a full-length protein with a predicted ... | 2001 | 11422507 |
| cloning, sequencing, tissue distribution, and heterologous expression of rat flavin-containing monooxygenase 3. | the sequence of rat fmo3 was obtained by rt-pcr and 5'/3' terminal extension. complete cdna was amplified, cloned, and sequenced. the cdna encodes a protein of 531 amino acids which contains the nadph- and fad-binding sites and a hydrophobic carboxyl terminus characteristic of fmos. this sequence is 81, 81, and 91% identical to sequences of human, rabbit, and mouse fmo3, respectively, and 60% identical to rat fmo1. rat fmo3 was expressed in escherichia coli. the recombinant protein and the nativ ... | 2001 | 11414682 |
| refocusing of b-cell responses following a single amino acid substitution in an antigen. | intranasal immunization of balb/c strain mice was carried out using baculovirus-derived human chorionic gonadotrophin (hcg) beta-chain, together with escherichia coli heat-labile enterotoxin. gonadotrophin-reactive immunoglobulin a (iga) was induced in a remote mucosal site, the lung, in addition to a systemic igg response. the extensive sequence homology with luteinizing hormone (lh) results in the production of lh cross-reactive antibodies when holo-hcg is used as an immunogen. in contrast to ... | 2001 | 11412304 |
| [expression in escherichia coli and immunogenicity of rotavirus vp7]. | rotaviruses are the single most important cause of severe diarrhea in young children all over the world. vp7 is the major outer capsid and is a primary candidate for inclusion in a subunit or recombinant vaccine. part of the vp7 gene containing all the three antigenic regions was expressed as a chimeric protein with glutathione s-transferase (gst) in e. coli. the chimeric protein, representing about 30% of the total protein of the recombinant-plasmid-carrying bacteria, reacted with polyclonal an ... | 2001 | 11411220 |
| effect of probiotics on enterocyte bacterial translocation in vitro. | enteral probiotics such as lactobacillus casei gg (lgg) have been used in the treatment of a variety of intestinal disorders in infants and children, including diarrhea, malabsorption, and clostridium difficile colitis. we have previously demonstrated that the probiotic bacterium lgg has an inhibitory effect on bacterial translocation (bt) in a neonatal rabbit model. however, this in-vivo model is limited for investigating the cellular and molecular mechanisms responsible for probiotic inhibitio ... | 2001 | 11409159 |
| visualization of secreted hrp and avr proteins along the hrp pilus during type iii secretion in erwinia amylovora and pseudomonas syringae. | pili are required for protein and/or dna transfer from bacteria to recipient plant or bacterial cells, based on genetic evidence. however, it has never been shown directly that the effector proteins or dna are localized along or inside the pili in situ. failure to visualize an association of effector proteins/dna with pili is the central issue in the debate regarding the exact function of pili in protein and dna transfer. in this study, a newly developed in situ immunogold labelling procedure en ... | 2001 | 11401717 |
| merozoite surface protein 8 of plasmodium falciparum contains two epidermal growth factor-like domains. | by motif searching of the unfinished sequences in the malaria genome sequencing project databases we have identified a novel egf-like domain-containing protein of plasmodium falciparum. the sequence lies within a single open reading frame of 1791 bp and is predicted to encode a polypeptide of 597 amino acids. there are hydrophobic regions at the extreme n- and c-termini, which could represent secretory signal peptide and gpi attachment sites, respectively. similar to msp1, there are two egf-like ... | 2001 | 11378201 |
| the kinetic and spectral characterization of the e. coli-expressed mammalian cyp4a7: cytochrome b5 effects vary with substrate. | the cyp4a gene subfamily is composed of a number of genes that encode cytochromes p450 from various species, including human, which catalyze the hydroxylation of various saturated and unsaturated fatty acids, including arachidonic acid and prostaglandins. cyp4a7, a fatty acid metabolizing cytochrome p450 from rabbit kidney, was expressed in e. coli by adding the first 10 codons of cyp17alpha producing final yields of 20 nmol/l in order to perform detailed kinetic and spectral studies. cyp4a7 met ... | 2001 | 11368012 |
| the polypeptide chain release factor erf1 specifically contacts the s(4)uga stop codon located in the a site of eukaryotic ribosomes. | it has been shown previously [brown, c.m. & tate, w.p. (1994) j. biol. chem. 269, 33164-33170.] that the polypeptide chain release factor rf2 involved in translation termination in prokaryotes was able to photocrossreact with mini-messenger rnas containing stop signals in which u was replaced by 4-thiouridine (s4u). here, using the same strategy we have monitored photocrosslinking to eukaryotic ribosomal components of 14-mer mrna in the presence of trna(f)(met), and 42-mer mrna in the presence o ... | 2001 | 11358506 |
| use of recombinant mitogillin for improved serodiagnosis of aspergillus fumigatus-associated diseases. | during human infection, aspergillus fumigatus secretes a 18-kda protein that can be detected as an immunodominant antigen in the urine of infected patients. recently, this protein was shown to be mitogillin, a ribotoxin that cleaves a single phosphodiester bond of the 29s rrna of eukaryotic ribosomes. we proved the immunogenic capacity of mitogillin in a rabbit animal model, indicating its usefulness as an antigen for serological diagnosis of invasive aspergillosis. the mitogillin gene from a. f ... | 2001 | 11325981 |
| development of a monoclonal sandwich elisa for the detection of animal and human escherichia coli o157 strains. | production of a monoclonal antibody (mab) to escherichia coli o157 to develop a rapid test using a sandwich elisa (selisa) format. | 2001 | 11309065 |
| protein kinase c phosphorylates nonmuscle myosin-ii heavy chain from drosophila but regulation of myosin function by this enzyme is not required for viability in flies. | conventional myosins (myosin-iis) generate forces for cell shape change and cell motility. myosin heavy chain phosphorylation regulates myosin function in simple eukaryotes and may also be important in metazoans. to investigate this regulation in a complex eukaryote, we purified the drosophila myosin-ii tail expressed in escherichia coli and showed that it was phosphorylated in vitro by protein kinase c(pkc) at serines 1936 and 1944, which are located in the nonhelical globular tail piece. these ... | 2001 | 11297427 |
| characterization of a heavy metal atpase from the apicomplexan cryptosporidium parvum. | p1-atpases are transporters which pump heavy metals across membranes, either to provide enzymes with essential cofactors or to remove excess, toxic metal cations from the cytosol. the first protist p1-atpase (cpatpase2) has been isolated from the apicomplexan cryptosporidium parvum, an opportunistic pathogen of aids patients. this single copy gene encodes 1260 amino acids (aa), predicting a protein of 144.7 kda. reverse transcription-polymerase chain reaction (rt-pcr) and western blot analysis c ... | 2001 | 11290416 |
| construction of a tagging system for subcellular localization of proteins encoded by open reading frames. | we have previously characterized a monoclonal antibody (sc1d7) that is directed to maltose-binding protein (mbp) of escherichia coli and other closely related enteric bacteria. sc1d7 does not cross-react with proteins in eucaryotes and appears to be a highly specific tool in immunochemical analyses. to better map the epitope, we took advantage of an available plasmid, pmal-c2, that encodes the e. coli mbp-coding sequence and constructed plasmids to express mbp fragments. a construct containing t ... | 2001 | 11287747 |
| bovine rotavirus nonstructural protein 4 produced by lactococcus lactis is antigenic and immunogenic. | rotavirus nonstructural protein 4 (nsp4) can induce diarrhea in mice. to get insight into the biological effects of nsp4, production of large quantities of this protein is necessary. we first tried to produce the protein in escherichia coli, but the nsp4 gene proved to be unstable. the capacity of the generally regarded as safe organism lactococcus lactis to produce nsp4 either intra- or extracellularly was then investigated by using the nisin-controlled expression system. production of recombin ... | 2001 | 11282586 |
| expression of rat liver long-chain acyl-coa synthetase and characterization of its role in the metabolism of r-ibuprofen and other fatty acid-like xenobiotics. | our investigations of fatty acid metabolism and epimerization of the 2-arylpropionic acid derivative, r-ibuprofen, resulted in the successful purification of an acyl-coa synthetase from rat liver microsomes that catalyzes the formation of both palmitoyl-coa and r-ibuprofenoyl-coa. to investigate whether r-ibuprofenoyl-coa synthetase and long-chain acyl-coa synthetase (lacs) are identical enzymes, we cloned the cdna from lacs into the pqe30 expression vector and transformed the construct into esc ... | 2001 | 11266649 |
| characterisation of the rna-dependent rna polymerase from rabbit hemorrhagic disease virus produced in escherichia coli. | all positive-strand rna viruses encode a rna-dependent rna polymerase which in most cases has been only identified on the basis of its sequence conservation. catalytic activity has been experimentally demonstrated in only a handful of these viral proteins, including that from rabbit hemorrhagic disease virus. studies from our laboratory have reported that rhdv rna polymerase produced in escherichia coli was enzymatically active showing poly(a)-dependent poly(u) polymerase as well as rna polymera ... | 2001 | 11266218 |
| characterization and functional analysis of the nucleotide binding fold in human peroxisomal atp binding cassette transporters. | the 70-kda peroxisomal membrane protein (pmp70) and the adrenoleukodystrophy protein (aldp) are half atp binding cassette (abc) transporters in the peroxisome membrane. mutations in the ald gene encoding aldp result in the x-linked neurodegenerative disorder adrenoleukodystrophy. plausible models exist to show a role for atp hydrolysis in peroxisomal abc transporter functions. here, we describe the first measurements of the rate of atp binding and hydrolysis by purified nucleotide binding fold ( ... | 2001 | 11248239 |
| expression of hbv pre s1 peptide in e. coli and product characterization. | hbv pre s1 sequence is supposed to play an important role in the infection of hbv. presence of pre s1/anti-pre s1 in serum has valuable clinical implications. in order to improve the study of pre s1, pre s1 sequence was overexpressed in e. coli as a fusion protein with mbp (maltose-binding protein), and anti-pre s1 antiserum was elicited in rabbits by pre s1-mbp purified by affinity chromatography. the recombinant plasmid constructed from pmal-cri expressed the 106aa pre s1 sequence at the c ter ... | 1997 | 11243097 |
| production of inflammatory cytokines by human gingival fibroblasts stimulated by cell-surface preparations of porphyromonas gingivalis. | porphyromonas gingivalis is a gram-negative rod associated with the progression of human periodontal disease. inflammatory cytokines are believed to be the major pathological mediators in periodontal diseases. we therefore investigated the productions of interleukin-1beta (il-1beta), interleukin-6 (il-6), interleukin-8 (il-8), and tumor necrosis factor-alpha (tnf-alpha) in human gingival fibroblasts treated with lipopolysaccharide, polysaccharide and outer-membrane proteins from p. gingivalis at ... | 2001 | 11240858 |
| an immunodominant membrane protein gene from the western x-disease phytoplasma is distinct from those of other phytoplasmas. | membrane proteins mediate several important processes, including attachment, in several mollicute species. phytoplasmas are non-culturable plant pathogenic mollicutes that are transmitted in a specific manner by certain phloem-feeding insect vectors. because it is likely that phytoplasma membrane proteins are involved with some aspect of the transmission process, their identification, isolation and characterization are important first steps in understanding phytoplasma transmission. a 32 kda imm ... | 2001 | 11238964 |
| active immunization with a detoxified escherichia coli j5 lipopolysaccharide group b meningococcal outer membrane protein complex vaccine protects animals from experimental sepsis. | the passive infusion of antibodies elicited in rabbits with a detoxified j5 lipopolysaccharide (lps)/group b meningococcal outer membrane protein complex vaccine protected neutropenic rats from heterologous lethal gram-negative bacterial infection. in this study, active immunization was studied in neutropenic rats infected with pseudomonas aeruginosa, in the presence or absence of ceftazidime therapy, and with klebsiella pneumoniae. this vaccine elicited a > 200-fold increase in anti-j5 lps anti ... | 2001 | 11237833 |
| human lens thioltransferase: cloning, purification, and function. | to clone the human lens thioltransferase (ttase) gene and to purify, characterize and study the possible function of the recombinant human lens thioltransferase (rhlt). | 2001 | 11222536 |
| properties of recombinant htra: an otitis media vaccine candidate antigen from non-typeable haemophilus influenzae. | non-encapsulated or non-typable haemophilus influenzae (nthi) is a major cause of middle ear infections in young children. htra has been identified as a vaccine candidate antigen from nthi; therefore physicochemical characterization of this antigen is important for vaccine development. recombinant nthi htra has been expressed in e. coli and shown to have serine protease activity. several mutant, recombinant htra proteins were expressed and purified to obtain suitable vaccine antigens lacking pro ... | 2000 | 11214237 |
| effect of natural antioxidants and apocynin on lps-induced endotoxemia in rabbit. | the objective of this study was to compare the prophylactic effects of the natural antioxidant from spinach (nao) and apocynin, on the hepatic oxidative stress and liver damage induced by lipopolysaccharide (lps). male new zealand rabbits were challenged with lps with or without 8 days of antioxidant pretreatment. pretreatment with nao, but not apocynin, significantly (p < 0.05) decreased the levels of hydroperoxides and malondialdehyde (mda) in the liver cytosolic fraction and the activity of n ... | 2000 | 11211238 |
| a 99tcm-labelled leukotriene b4 receptor antagonist for scintigraphic detection of infection in rabbits. | in a search for a rapid and accurate imaging agent for scintigraphic detection of infection and inflammation, an ltb4 receptor antagonist, 99tcm-rp517, which contains the hydrazino nicotinamide moiety, has been developed recently. to study the in vivo behaviour of 99tcm-rp517, rabbits with escherichia coli infection were injected intravenously with 99tcm-rp517. gamma camera images were obtained and ex vivo bio-distribution was determined at several hours post-injection (p.i.). in a separate set ... | 2000 | 11192710 |
| molecular cloning and functional expression of bottle-nosed dolphin (tursiops truncatus) interleukin-1 receptor antagonist. | the bottle-nosed dolphin (tursiops truncatus) interleukin-1 receptor antagonist il-1ra cdna was cloned from mitogen-stimulated peripheral blood mononuclear cells (pbmc) rna utilizing the reverse transcription-polymerase chain reaction (rt-pcr). the sequence of this cdna showed that dolphin il-1ra clones contained open reading frames encoding 177 amino acids. comparison of the deduced amino acids showed that dolphin il-1ra sequence shared 87.6, 77.9, 77.4, 77.4, 76.4, and 75.8% similarity with th ... | 2001 | 11182153 |
| recombinant neisseria meningitidis transferrin binding protein a protects against experimental meningococcal infection. | to better characterize the vaccine potential of neisseria meningitidis transferrin binding proteins (tbps), we have overexpressed tbpa and tbpb from neisseria meningitidis isolate k454 in escherichia coli. the ability to bind human transferrin was retained by both recombinant proteins, enabling purification by affinity chromotography. the recombinant tbps were evaluated individually and in combination in a mouse intraperitoneal-infection model to determine their ability to protect against mening ... | 2001 | 11179327 |
| bacterial expression of an immunologically reactive pcv2 orf2 fusion protein. | the entire coding region of open reading frame 2 (orf2) of porcine circovirus 2 (pcv2) was linked to the 3'-end of the maltose-binding protein (mbp)-his(8)-tag gene. the fusion protein was expressed as soluble form after induction by isopropylthio-beta-d-galactoside. mbp-his(8)-orf2 was purified to homogeneity by immobilized metal affinity chromatography based on the interaction of the polyhistidine-tag with metal ions. expression could represent 1% of the total protein in escherichia coli, allo ... | 2001 | 11162395 |
| a new expression cloning strategy for isolation of substrate-specific kinases by using phosphorylation site-specific antibody. | signal transduction from cell surface receptors to the nucleus is regulated in most part by protein phosphorylation. for the purpose of identification of kinases which play an important role at a particular phosphorylation step in a series of signal transduction pathways, we have developed a new expression-screening method using a phosphorylation site specific antibody and a vector encoding substrate polypeptide. we have applied this method for screening kinases which phosphorylate stat3 at seri ... | 2001 | 11150545 |
| impairment of cardiac beta-adrenoceptor cellular signaling by decreased expression of g(s alpha) in septic rabbits. | abnormalities in the beta-adrenergic control of cardiac function play a role in the pathogenesis of several disease states. because circulatory failure in patients with septic shock is known to be less responsive to catecholamines, we investigated whether the beta-adrenoceptor-linked signal transduction mechanisms are altered in the heart of a septic animal model | 2000 | 11149442 |
| cleavage of cyclin a at r70/r71 by the bacterial protease ompt. | previous work has shown that cyclin a can be cleaved at arg-70/arg-71 by a proteolytic activity present in an in vitro-coupled transcription/translation system by using rabbit reticulocyte lysate programmed by plasmid dna encoding p27(kip1), a cyclin-dependent kinase inhibitor, but not by plasmid dnas encoding other cyclin-dependent kinases inhibitors. here we report that cyclin a is also cleaved by translation product programmed by plasmid dna encoding cyclin b. several findings indicate that t ... | 2001 | 11136238 |
| immunoprophylactic potential of cloned shiga toxin 2 b subunit. | the shiga toxins stx1 and stx2 contribute to the development of enterohemorrhagic o157:h7 escherichia coli-mediated colitis and hemolytic-uremic syndrome in humans. the stx2 b subunit, which binds to globotriaosylceramide (gb3) receptors on target cells, was cloned. this involved replacing the stx2 b subunit leader peptide nucleotide sequences with those from the stx1 b subunit. the construct was expressed in the topp3 e. coli strain. the stx2 b subunits from this strain assembled into a pentame ... | 2001 | 11133375 |
| three-dimensional structure of rk-1: a novel alpha-defensin peptide. | nmr spectroscopy and simulated annealing calculations have been used to determine the three-dimensional structure of rk-1, an antimicrobial peptide from rabbit kidney recently discovered from homology screening based on the distinctive physicochemical properties of the corticostatins/defensins. rk-1 consists of 32 residues, including six cysteines arranged into three disulfide bonds. it exhibits antimicrobial activity against escherichia coli and activates ca(2+) channels in vitro. through its p ... | 2000 | 11123900 |
| an investigation into the pathogenic properties of escherichia coli strains blr, bl21, dh5alpha and eq1. | to examine escherichia coli strains eq1, dh5alpha, blr and bl21 for known pathogenic mechanisms. | 2000 | 11123478 |
| enterotoxigenic escherichia coli tiba glycoprotein adheres to human intestine epithelial cells. | enterotoxigenic escherichia coli (etec) is capable of invading epithelial cell lines derived from the human ileum and colon. two separate invasion loci (tia and tib) that direct noninvasive e. coli strains to adhere to and invade cultured human intestine epithelial cells have previously been isolated from the classical etec strain h10407. the tib locus directs the synthesis of tiba, a 104-kda outer membrane glycoprotein. synthesis of tiba is directly correlated with the adherence and invasion ph ... | 2001 | 11119488 |
| the asp-rich region at the carboxyl-terminus of calsequestrin binds to ca(2+) and interacts with triadin. | calsequestrin (csq) is a high capacity ca(2+) binding protein in the junctional sarcoplasmic reticulum of striated muscles, and has been shown to regulate the ryanodine receptor (ryr) through triadin and junctin. in order to identify the functional roles of specific regions on csq, several csq deletion mutants were prepared by molecular cloning and escherichia coli expression. 45ca(2+) overlay assay using a native gel system revealed that the major ca(2+) binding motif of csq resides in the asp- ... | 2000 | 11113462 |
| a single endotoxin injection in the rabbit causes prolonged blood vessel dysfunction and a procoagulant state. | to determine the duration of vascular blood vessel dysfunction and coagulation abnormalities after administration of endotoxin in a nonlethal septic rabbit model. | 2000 | 11098972 |
| histologic inflammation in the maternal and fetal compartments in a rabbit model of acute intra-amniotic infection. | this study was undertaken to determine the course of acute inflammation in the maternal and fetal compartments during experimentally induced ascending intra-amniotic infection. | 2000 | 11084546 |
| epithelial cell adherence mediated by the enterotoxigenic escherichia coli tia protein. | in vitro studies have shown that enterotoxigenic escherichia coli (etec) strains are capable of invading cultured epithelial cells derived from the human ileum and colon. two separate invasion loci (tia and tib) have previously been isolated from the classical etec strain h10407. the tia locus has been shown to direct the synthesis of tia, a 25-kda outer membrane protein. tia is sufficient to confer the adherence and invasion phenotypes on laboratory stains of e. coli, suggesting that this prote ... | 2000 | 11083770 |
| selecting and evolving functional proteins in vitro by ribosome display. | 2000 | 11075357 | |
| contribution of defined amino acid residues to the immunogenicity of recombinant escherichia coli heat-stable enterotoxin fusion proteins. | we investigated whether the toxicity-associated receptor-binding domain of the non-immunogenic escherichia coli heat-stable enterotoxin (sth) as a fusion with a carrier protein and the inclusion of an appropriate spacer are critical factors for eliciting antibody responses against the native toxin. the immunological properties of three toxic and one non-toxic fusion proteins, consisting of sth n-terminally joined to the c-terminus of the major subunit clpg of e. coli cs31a fimbriae, were compare ... | 2000 | 11064199 |
| effect of l-arginine on endothelial injury and hemostasis in rabbit endotoxin shock. | to investigate whether impaired endothelial function was related to alteration of nitric oxide (no) formation during endotoxic shock, we studied the effects of supplementation of l-arginine (l-arg), d-arginine (d-arg), and n(g)-nitro-l-arginine methyl ester (l-name), on endothelial function and structure in a rabbit model. endotoxic shock was induced by a single lipopolysaccharide bolus (0.5 mg/kg i.v., escherichia coli endotoxin). coagulation factors and expression of monocyte tissue factor wer ... | 2000 | 11053330 |
| functional recombinant rabbit muscle phosphoglucomutase from escherichia coli. | the gene coding for phosphoglucomutase (pgm) from oryctolagus cuniculus (rabbit) has been expressed in escherichia coli under a t7 expression system with a his-tag. about half of the expressed pgm protein was present in inclusion bodies, but this protein was inactive when solubilized. the protein in the soluble cell fraction was isolated and purified in one step on a ni-nta column. the eluate from this column was adjusted to 95% saturated ammonium sulfate, and the resulting protein precipitate w ... | 2000 | 11035960 |
| tracing the origin of the ractk1 k(+) channel. | potassium secretion by the kidney is vital for the maintenance of k(+) homeostasis. ractk1, a putative inwardly rectifying potassium channel cloned from cultured rabbit collecting duct cells, has been proposed to play a role in this process. however, the lack of homology with any other cloned potassium channel and the inability to reproduce the results across different laboratories has brought into question the existence of ractk1. recently, it has been suggested that ractk1 is a contamination f ... | 2000 | 11027655 |
| development of eia for detection of chlamydia trachomatis in genital specimens. | a double antibody sandwich enzyme immunoassay (eia) for chlamydial antigen detection was developed using a monoclonal antibody against lipopolysaccharide (lps) of chlamydia trachomatis as a coating antibody. polyclonal rabbit antiserum against partially purified antigen from elementary body (eb) antibody and horse-radish peroxidase conjugated goat anti-rabbit antibody were used as the primary and secondary antibody respectively. the developed eia could detect protein of partially purified eb at ... | 2000 | 11023074 |
| pet toxin from enteroaggregative escherichia coli produces cellular damage associated with fodrin disruption. | pet toxin is a serine protease from enteroaggregative escherichia coli which has been described as causing enterotoxic and cytotoxic effects. in this paper we show that pet produces spectrin and fodrin (nonerythroid spectrin) disruption. using purified erythrocyte membranes treated with pet toxin, we observed degradation of alpha- and beta-spectrin chains; this effect was dose and time dependent, and a 120-kda protein fraction was observed as a breakdown product. spectrin degradation and product ... | 2000 | 10992503 |
| genetic characterization of dna region containing the trh and ure genes of vibrio parahaemolyticus. | we have demonstrated that possession of the gene for thermostable direct hemolysin-related hemolysin (trh) coincides with the presence of the urease gene among clinical vibrio parahaemolyticus strains and that the location of the two genes are in close proximity on the chromosome. here, we cloned and sequenced the 15,754-bp dna region containing the trh gene and the gene cluster for urease production from the chromosome of clinical v. parahaemolyticus (th3996). we found 16 open reading frames (o ... | 2000 | 10992480 |
| is1414, an escherichia coli insertion sequence with a heat-stable enterotoxin gene embedded in a transposase-like gene. | enteroaggregative escherichia coli (eaec) heat-stable enterotoxin 1 (east1) was originally discovered in eaec but has also been associated with enterotoxigenic e. coli (etec). multiple genomic restriction fragments from each of three etec strains of human origin showed homology with an east1 gene probe. a single hybridizing fragment was detected on the plasmid of etec strain 27d that also encodes heat-stable enterotoxin ib and colonization factor antigen i. we isolated and characterized this fra ... | 2000 | 10992475 |
| substrate specificity analysis of microbial transglutaminase using proteinaceous protease inhibitors as natural model substrates. | the substrate specificity of microbial transglutaminase (mtg) from streptomyces mobaraensis (formerly categorized streptoverticillium) was studied using a streptomyces proteinaceous protease inhibitor, sti2, as a model amine-donor substrate. chemical modification and mutational analysis to address the substrate requirements for mtg were carried out around the putative reactive site region of sti2 on the basis of the highly refined tertiary structure and the solvent accessibility index of strepto ... | 2000 | 10965040 |
| a flexible single-step detection of blotted antigen using a fusion protein between protein a and green fluorescent protein. | a green fluorescent protein mutant (s147p gfp) was fused with protein a and expressed in escherichia coli. this fusion protein (pa-gfp147) was used in immunoblotting studies as a new detection system, designated as "flexible single-step detection (fssd)". in fssd, the detection of blotted antigen was done in one step, and the antigen-antibody reaction can be monitored by uv-irradiation in real time. the reaction time, therefore, can be flexibly controlled by monitoring the green fluorescence. | 2000 | 10945281 |
| cloning, sequencing, and expression of the tulip bulb chitinase-1 cdna. | a cdna encoding tulip bulb chitinase-1 (tbc-1) was cloned using a combination of immunoscreening from a lambda zap cdna library with anti-tbc-1 antiserum and the 5' rapid amplification of cdna end (race) method, and sequenced. the cdna consists of 1,106 nucleotides and included an open reading frame encoding a polypeptide of 314 amino acids. comparison of the deduced amino acid sequence and the determined protein sequence indicated the presence of a signal peptide and an extra peptide composed o ... | 2000 | 10945255 |
| mechanisms of action of rabbit cap18 on monolayers and liposomes made from endotoxins or phospholipids. | we have investigated the mechanism of action of the cationic antimicrobial protein (18 kda) cap18 on liposomes and monolayers made from phospholipids and enterobacterial lipopolysaccharides (lps). cap18 intercalates into lipid matrices composed of lps from sensitive strains, weaker into those made of lps from a resistant strain (proteus mirabilis strain r45) or negatively charged phospholipids, but not into those composed of neutral phosphatidylcholine. from the combination of data obtained with ... | 2000 | 10931974 |
| functional characterization of green fluorescent protein-profilin fusion proteins. | to clarify the role of profilins in cells, fusion proteins constructed with green fluorescent protein (gfp) should be extremely helpful. as profilins are considerably smaller than the gfp fusion partner (14-17 kda compared with 27 kda, respectively), we characterized the fusion proteins in vitro, to ascertain their biological function. we fused mouse profilin i and ii to either the c-terminus or n-terminus of gfp. these fusion proteins were expressed in escherichia coli and affinity-purified on ... | 2000 | 10931210 |
| [effects of specific removal of circulating tumor necrosis factor-alpha by immunoadsorption on nitric oxide in endotoxin shock]. | to evaluate the effects of specific removal of circulating tnf-alpha by immunoadsorption on nitric oxide (no) in endotoxin shock. | 1998 | 10923398 |
| molecular cloning and characterization of a unique 60 kda/72 kda antigen gene encoding enzyme i of the phosphoenolpyruvate: sugar phosphotransferase system (pts) of mycoplasma hyopneumoniae. | the recombinant clone expressing a 60 kda (p60) antigen was isolated from escherichia coli by screening a lambda embl3 genomic library using rabbit produced antiserum against mycoplasma hyopneumoniae. sequence analysis revealed that an interrupted (by a uga codon) open reading frame coding for a 72 kda protein (p72) may contain the p60 antigen gene. western blot analysis with an anti-p60 monospecific antibody confirmed the presence of a p72 antigen from the total protein of m. hyopneumoniae, and ... | 2000 | 10920262 |