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At least 1,531 records · Page 85Linked to original sources
PubMed2002

Identification of the pepper SAR8.2 gene as a molecular marker for pathogen infection, abiotic elicitors and environmental stresses in Capsicum annuum.

Pepper ( Capsicum annuum L.) SAR8.2 genes, designated CASAR82A, B and C, which are induced by all the biotic and abiotic stresses, were isolated from a pepper cDNA library constructed with the mRNAs from pepper plants infected with Xanthomonas campestris pv. vesicatoria. The pepper CASAR82A, B and C gene products, which are very similar to each other in amino acid sequences, have 43-50% homology with those of tobacco SAR8.2 genes. The CASAR8.2 genes were not constitutively expressed in any of the organs of healthy pepper plants. In contrast, the CASAR82A gene was locally or systemically induced in pepper plants infected by X. campestris pv. vesicatoria, Colletotrichum coccodes or Phytophthora capsici. Strong induction of the CASAR82A gene also was found in pepper leaves treated with ethylene, methyl jasmonate, indole-3-acetic acid, abscisic acid, salicylic acid, benzothiadiazole, DL-beta- n-amino butyric acid or hydrogen peroxide. Interestingly, the transcription of the CASAR82A gene was rapidly triggered by high salinity, drought or low-temperature stresses, but not by mechanical wounding. In situ hybridization results revealed that the CASAR82A mRNAs were localized in phloem and epidermal cells of pepper leaf and stem tissues infected by C. coccodes and P. capsici, or treated with salicylic acid. These results thus suggest that pepper SAR8.2 genes may be valuable as a molecular marker for the detection of various pathogen infections, abiotic elicitors and environmental stresses.

Adaptation, Physiological↗
PubMed2001

Identification of two highly divergent catalase genes in the fungal tomato pathogen, Cladosporium fulvum.

Catalases of pathogenic micro-organisms have attracted attention as potential virulence factors. Homology-based screens were performed to identify catalase genes in the fungal tomato pathogen Cladosporium fulvum. Two highly divergent genes, Cat1 and Cat2, were isolated and characterized. Cat1 codes for a putative 566-amino-acid catalase subunit and belongs to the gene family that also encodes the mainly peroxisome-localized catalases of animal and yeast species. Cat2 codes for a putative catalase subunit of 745 amino acids and belongs to a different gene family coding for the large-subunit catalases similar to ones found in bacteria and filamentous fungi. Neither catalase had an obvious secretory signal sequence. A search for an extracellular catalase was unproductive. The Cat1 and Cat2 genes showed differential expression, with the Cat1 mRNA preferentially accumulating in spores and the Cat2 mRNA preferentially accumulating in response to external H(2)O(2). With Cat2-deleted strains, activity of the Cat2 gene product (CAT2) was identified among four proteins with catalase activity separated on non-denaturing gels. The CAT2 activity represented a minor fraction of the catalase activity in spores and H(2)O(2)-stressed mycelium, and no phenotype was observed for Cat2-deleted strains, which showed a normal response to H(2)O(2) treatment. These results indicate the existence of a complex catalase system in C. fulvum, with regard to both the structure and regulation of the genes involved. In addition, efficient C. fulvum gene-replacement technology has been established.

Amino Acid Sequence↗
PubMed1999

Identification and characterization of a ran gene promoter in the protozoan pathogen Giardia lamblia.

The promoter elements that regulate transcription initiation in Giardia lamblia are poorly understood. In this report, the promoter of the Giardia ran gene was studied using a luciferase expression plasmid pRANluc+ to monitor transcription efficiency. An AT-rich sequence spanning -51/-20 relative to the translation start site of the ran gene was identified and was found to be required for efficient luciferase expression by deletion and mutation mapping of pRANluc+. The -51/-20 sequence was also sufficient for promoter activity as revealed from studies on a 32-base pair synthetic promoter derived from this region. Deletion mapping of the synthetic promoter revealed two minimal promoter elements, -51/-42 and -30/-20, sufficient for 6- and 30-fold luciferase expression above background, respectively. The transcription start sites on luc+ messenger RNA were determined by the position of the synthetic promoter in the luciferase expression plasmids as shown by primer extension experiments. Results from electrophoretic mobility shift assays revealed multiple DNA-protein complexes upon binding of nuclear proteins with either DNA strand but not the double-stranded DNA derived from the ran promoter. Our results delineate the first promoter sequence of the Giardia gene (ran), which provides an excellent model for future studies on transcription regulation in this protozoan parasite.

Animals↗
PubMed2006

Genomic island identification in Vibrio vulnificus reveals significant genome plasticity in this human pathogen.

UNLABELLED: Genomic islands (GIs) are large chromosomal regions present in a subset of bacterial strains that increase the fitness of the organism under specific conditions. We compared the complete genome sequences of two Vibrio vulnificus strains YJ016 and CMCP6 and identified 14 regions (ranging in size from 14 to 117 kb), which had the characteristics of GIs. Bioinformatic analysis of these 14 GI regions identified the presence of phage-like integrase genes, aberrant GC content and genome signature (dinucleotide frequency) within each GI compared with the core genome indicating that these regions were acquired from an anomalous source. We examined the distribution of the nine GIs from strain YJ016 among 27 V. vulnificus isolates and found that most GIs were absent from the majority of these isolates. The chromosomal insertion sites of three GIs were adjacent to tRNA sites, which contained novel horizontally acquired DNA in all six available sequenced Vibrionaceae genomes. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Adaptation, Physiological↗
PubMed2005

Identification of arabidopsis loci required for susceptibility to the downy mildew pathogen Hyaloperonospora parasitica.

Plants are susceptible to a limited number of pathogens. Most infections fail due to active defense or absence of compatibility. Many components of the plant's surveillance system and defense arsenal have been identified in the last decades. However, knowledge is limited on compatibility; in particular, the role of plant factors in the infection process. To gain insight into these processes, we have initiated an Arabidopsis thaliana mutant screen for reduced susceptibility to the downy mildew pathogen Hyaloperonospora parasitica. Ethyl methane sulfonate (EMS) mutants were generated in the highly susceptible Arabidopsis line Ler eds1-2. Eight downy mildew-resistant (dmr) mutants were analyzed in detail, corresponding to six different loci. Microscopic analysis showed that, in all mutants, H. parasitica growth was severely reduced. Resistance of dmr3, dmr4, and dmr5 was associated with constitutive expression of PR-1. Furthermore, dmr3 and dmr4, but not dmr5, also were resistant to Pseudomonas syringae and Golovinomyces orontii, respectively. However, enhanced activation of plant defense was not observed in dmr1, dmr2, and dmr6. We postulate that, in these susceptibility mutants, cellular processes are disrupted which are required for H. parasitica infection. This interesting new set of mutants provides a basis to elucidate the molecular processes underlying susceptibility to downy mildew in Arabidopsis.

Arabidopsis↗
PubMed1994

Identification and molecular cloning of four cysteine proteinase genes from the pathogenic protozoon Trichomonas vaginalis.

The parasitic protozoon Trichomonas vaginalis produces multiple forms of cysteine proteinase (CP). The molecular basis for this has now been examined by cloning DNA fragments encoding CPs. Using generic degenerate oligonucleotide primers based on two well-conserved regions within the central region of all eukaryotic CPs, several polymerase chain reaction fragments were isolated from T. vaginalis genomic DNA and shown to encode different CPs. One fragment with a well-represented sequence was used as a general probe to screen a T. vaginalis cDNA library at moderate stringency and five different cDNA clones were isolated. Preliminary sequencing showed that they encoded similar but distinct CPs. In the process of confirming the 5' end of one of these cDNA clones using RACE-PCR (rapid amplification of cDNA 5' ends-polymerase chain reaction), an additional sequence encoding a different CP was identified. The corresponding clone (TvCP3) and the three longest clones from the library screen (TvCP1, TvCP2 and TvCP4) were characterized further. TvCP1 and TvCP2 were full-length and TvCP3 and TvCP4 were apparently slightly less than full-length. Comparison of the predicted amino acid sequences of the four clones showed that TvCP1 and TvCP4 are related (72% identity). TvCP2 is closer to TvCP1 (60%) and TvCP4 (65%) than is TvCP3, which has 53%, 59% and 56% identity to TvCP1, TvCP2 and TvCP4, respectively. Comparison with the sequences of other known CPs indicated that the T. vaginalis gene products all belong to the cathepsin L/cathepsin H/papain branch of the papain superfamily. The TvCP1, TvCP2 and TvCP4 sequences are related (38-45% identity) to those of CP2 of Dictyostelium discoideum, human cathepsin L, three CPs from lobster and CPs from black gram, oilseed rape and rice (oryzains alpha and beta). TvCP3 shows less identity to the other eukaryotic CPs but is most similar to D. discoideum CP2 (38%). The four predicted amino acid sequences share some features distinct from the majority of CPs, which suggests they might have had a common evolutionary origin. The most striking feature of sequences TvCP1, TvCP2 and TvCP3 is the apparent lack of a pre-sequence (signal sequence) for TvCP1 and very short pre-sequences for TvCP2 and TvCP3. Southern analysis indicated that the organization of the genes corresponding to the TvCP cDNAs differs. The TvCP1, TvCP2 and TvCP3 genes are single-copy, whereas the TvCP4 gene appeared to be multiple-copy.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗
PubMed1999

Identification of a mating type-like locus in the asexual pathogenic yeast Candida albicans.

Candida albicans, the most prevalent fungal pathogen in humans, is thought to lack a sexual cycle. A set of C. albicans genes has been identified that corresponds to the master sexual cycle regulators a1, alpha1, and alpha2 of the Saccharomyces cerevisiae mating-type (MAT) locus. The C. albicans genes are arranged in a way that suggests that these genes are part of a mating type-like locus that is similar to the mating-type loci of other fungi. In addition to the transcriptional regulators a1, alpha1, and alpha2, the C. albicans mating type-like locus contains several genes not seen in other fungal MAT loci, including those encoding proteins similar to poly(A) polymerases, oxysterol binding proteins, and phosphatidylinositol kinases.

1-Phosphatidylinositol 4-Kinase↗
PubMed1992

Identification and cloning of a tetracycline resistance gene from the fish pathogen Vibrio salmonicida.

Vibrio salmonicida is the causative agent of cold-water vibriosis in farmed Atlantic salmon. We cloned a 6.9-kb PstI fragment from the 170-MDa plasmid (pRVS1) containing a tetracycline resistance determinant. A subcloned 1.96-kb HindIII fragment was found to mediate the tetracycline resistance. The 2.5-kb ClaI-PvuI fragment carrying the Tet E determinant from Escherichia coli hybridized under stringent conditions with the cloned 1.96-kb HindIII fragment from V. salmonicida. The 1.96-kb HindIII fragment codes for a protein of 26.5 kDa which represents a candidate for the structural TetA protein of the class E determinant. Deletion of a 0.28-kb BalI fragment from the middle of the HindIII fragment resulted in the loss of tetracycline resistance. We were able to show that when E. coli carries the cloned 6.9-kb PstI fragment, expression of tetracycline resistance is regulated by the concentration of tetracycline in the medium. In contrast, tetracycline resistance was constitutively expressed in the E. coli isolate carrying the 1.96-kb HindIII fragment. The tetracycline resistance gene isolated from the 170-MDa R plasmid of the marine fish pathogen V. salmonicida was characterized and shown to be a class E determinant.

Escherichia coli↗
PubMed2006

Identification of low-temperature-regulated genes in the fire blight pathogen Erwinia amylovora.

Genes involved in pathogenicity of several plant pathogens were shown to be induced at relatively cold temperatures. Loci from the fire blight pathogen Erwinia amylovora (Burrill) induced at 18 degrees C were identified using the miniTn5 transposon that contains the promoterless reporter gene gusA coding for beta-glucuronidase (GUS). Certain mutants (2.7%) expressed GUS predominantly at 18 degrees C on minimal medium plates, indicating that the transposon had been inserted downstream of a putatively thermoregulated promoter. Those mutants were further screened with a quantitative GUS fluorometric assay. A total of 21 mutants were selected: 19 mutants had a transposon insertion in temperature-dependent genetic loci, with a 2.2- to 6.3-fold induction of gusA gene expression at 18 degrees C, and two mutants with impaired growth at 18 degrees C. Some of these genetic loci encoded (i) proteins implicated in flagella biosynthesis, biotin biosynthesis, multi-drug efflux, and type II secretion protein, and (ii) proteins of unknown function.

Bacterial Proteins↗
PubMed1992

Identification of the defect in lipophosphoglycan biosynthesis in a non-pathogenic strain of Leishmania major.

The major macromolecule on the surface of the protozoan parasite, Leishmania major, is a complex lipophosphoglycan (LPG), which is anchored to the plasma membrane by an inositol-containing phospholipid. A defect in LPG biosynthesis is thought to be responsible for the avirulence of the L. major strain LRC L119 in mice. In order to identify the nature of this defect we have characterized two truncated forms of LPG, which are accumulated in this strain, by one- and two-dimensional 500-MHz 1H NMR spectroscopy, two-dimensional heteronuclear 1H-31P NMR spectroscopy, methylation analysis, and exoglycosidase digestions. The structures of these glycoinositolphospholipids, termed GIPL-4 and -6, are as follows: [formula: see text] The glycan moieties of GIPL-4 and -6 are identical to the anchor region of LPG, which is also substituted with a Glc-1-PO4 residue in approximately 60% of the structures. However, instead of being capped with chains of phosphorylated oligosaccharide repeat units, both glycan moieties terminate in Man alpha 1-PO4, suggesting that the defect in LPG biosynthesis is in the transfer of galactose to this residue to form the disaccharide backbone of the first repeat unit. These results indicate that the phosphoglycan moiety of LPG is essential for intracellular survival of the parasite and have implications for LPG biosynthesis.

Animals↗
PubMed1999

Identification of Acinetobacter baumannii strains with monoclonal antibodies against the O antigens of their lipopolysaccharides.

Despite the emergence of Acinetobacter baumannii strains as nosocomial pathogens, simple methods for their phenotypic identification are still unavailable. Murine monoclonal antibodies specific for the O-polysaccharide moiety of the lipopolysaccharide (LPS) of two A. baumannii strains were obtained after immunization with heat-killed bacteria. The monoclonal antibodies were characterized by enzyme immunoassay and by Western and dot blot analyses and were investigated for their potential use for the identification of A. baumannii strains. The antibodies reacted with 46 of the 80 A. baumannii clinical isolates that were investigated, and reactivity was observed with 11 of 14 strains which were isolated during outbreaks in different northwestern European cities; no reactivity was observed with Acinetobacter strains of other genomic species, including the closely related genomic species 1 (Acinetobacter calcoaceticus), 3, and 13 sensu Tjernberg and Ursing, or with other gram-negative bacterial strains. The results show that O-antigen-specific monoclonal antibodies such as the ones described are convenient reagents which can be used to identify Acinetobacter strains in clinical and research laboratories.

Acinetobacter↗
PubMed2026

Whole-genome prediction of bacterial pathogenic capacity on novel bacteria using protein language models with PathogenFinder2.

MOTIVATION: Infectious diseases continue to be a leading cause of mortality and pose a significant global health threat. Thus, the development of tools for surveillance and early detection of emerging pathogens is needed. RESULTS: We introduce PathogenFinder2, a novel, alignment-free, taxonomy-agnostic model for predicting bacterial pathogenic capacity in humans using protein language models. It outperforms previous methods, particularly for novel taxa, and provides interpretable outputs by highlighting proteins most relevant to pathogenic potential. These insights aid the identification of virulence factors, vaccine targets, and infection-related metabolic pathways. Furthermore, we introduce the Bacterial Pathogenic Capacity Landscape, which reveals patterns linked to host condition, infection site, microbial antagonism, and environmental origin. AVAILABILITY: The model is freely available online at https://genepi.dk/pathogenfinder2, or as a standalone program (https://github.com/genomicepidemiology/PathogenFinder2).

Genome, Bacterial↗
PubMedDate not supplied

New methods for identification of obligate anaerobes.

Anaerobes are identified in the clinical laboratory to provide a microbiologic diagnosis for the individual patient as well as information on pathogenicity and epidemiology. The level of identification required in each context should be discussed and agreement should be reached. New methods of identification available, or likely to become available, include nucleic acid and immunologic probes, protein electrophoresis, peptidoglycan analysis, detection of volatile metabolic products, and detection of enzymes. The last of these has been widely adopted for the production of commercially available identification kits. All the methods require critical assessment before they are introduced into routine service.

Bacteria, Anaerobic↗
PubMed2006

Diagnosis of American foulbrood in honey bees: a synthesis and proposed analytical protocols.

Worldwide, American foulbrood (AFB) is the most devastating bacterial disease of the honey bee (Apis mellifera). Because the distinction between AFB and powdery scale disease is no longer considered valid, the pathogenic agent has recently been reclassified as one species Paenibacillus larvae, eliminating the subspecies designations Paenibacillus larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens. The creamy or dark brown, glue-like larval remains of infected larvae continue to provide the most obvious clinical symptom of AFB, although it is not conclusive. Several sensitive and selective culture media are available for isolation of this spore-forming bacterium, with the type of samples that may be utilized for detection of the organism being further expanded. PCR methods for identification and genotyping of the pathogen have now been extensively developed. Nevertheless, biochemical profiling, bacteriophage sensitivity, immunotechniques and microscopy of suspect bacterial strains are entirely adequate for routine identification purposes.

Animals↗
PubMed1985

Unique features of group B streptococcal arthritis in adults.

Group B streptococci have rarely been reported to cause serious infection in adults and even less frequently to result in septic arthritis. We reviewed our clinical and radiologic experience with septic arthritis and uncovered five cases of infectious arthritis caused by group B streptococci. Unlike previously described patients with monoarthritis and complete recovery, our patients displayed polyarticular involvement. The infection in some cases was aggressive, resulting in destruction of multiple joints. Therefore, the series supports the view that this uncommon pathogen may produce an aggressive polyarthritis with the potential for serious functional damage and permanent morbidity. Culture and identification of a group B streptococcal pathogen and prompt institution of therapy can help avoid these complications.

Arthritis, Infectious↗
PubMed2002

Microbiologic findings and risk factors for antimicrobial resistance at myringotomy for tympanostomy tube placement--a prospective study of 601 children in Toronto.

CONTEXT: There is limited information on the identity and antibiotic susceptibility of bacterial pathogens in children with chronic otitis media whose repeated antibiotic use may place them at increased risk of antibiotic-resistant bacteria. OBJECTIVE: To determine, at myringotomy for tympanostomy tube placement, (1) the prevalence of bacteria, (2) the extent and patterns of antibiotic resistance, and (3) the risk factors associated with the presence and resistant status of pathogens. DESIGN: Prospective, multi-site, cohort study. SETTING AND PATIENTS: Children undergoing myringotomy for tympanostomy tube placement between November 1, 1999 and March 31, 2000 in seven hospitals in Toronto, Ontario, were identified. If fluid was present, aspirates were submitted for bacteriologic testing. A follow-up telephone questionnaire was administered to patient caregivers in order to identify risk factors for the presence of (1) culturable pathogens and (2) resistant pathogens. MAIN OUTCOME MEASURES: The identification and prevalence of bacteria cultured from the middle ears of subjects, and the degree of nonsusceptibility to commonly prescribed antibiotics. RESULTS: Among 601 patients (mean age 3.9 years, 60.7% male), both a telephone interview (n=544) and an ear specimen (n=527) were obtained for 478. Pathogens were found in middle ear effusions of 37% of the children in the study; including at least one 'definite' pathogen in 189 children (31.4%), and a further 32 children (5.3%) with at least one 'possible' pathogen. Definite pathogens included Haemophilus influenzae in 17% of the children, followed by Moraxella catarrhalis (9%) and Streptococcus pneumoniae (6%); ampicillin nonsusceptibility was found in 40, 100 and 24%, respectively. Overall, 123 children (20.5%) were found to have definite pathogens with resistance to ampicillin/penicillin, trimethoprim-sulfamethoxazole, or clarithromycin/erythromycin. Patient characteristics included premature birth and/or long length of stay in the nursery (23%), first infection before the age of 6 months (26%), put to bed with a bottle (28%), household smoker (34%), in out-of-home child care (38%), history of eczema, bronchiolitis and/or asthma (39%), and use of pacifiers (40%). Household characteristics were smoking (34%), married/common law parents (85%), and 60% had completed college or university; in 26% both parents were born outside of Canada; 73% of children were Caucasian. Of the 75% who responded to the question regarding income, 42% had household income over $60,000 (CAN). Risk factors for the presence of a pathogen and for a resistant pathogen in multivariate analysis included younger age, lower maternal education, day care centre attendance, no previous adenoidectomy and bilateral, primarily winter infections as well as amoxicillin use in the previous 6 months. CONCLUSION: Modifiable risk factors for otitis media including household smoking and pacifier use are present in many children undergoing tympanostomy tube placement; child care centre attendees are over-represented. Multiple antibiotic courses were commonly prescribed prior to surgery. H. influenzae and M. catarrhalis are important pathogens and therapy in clinical failures should be directed against them. The 7-valent protein conjugate polysaccharide vaccine (Prevnar) would have covered 73% of the serotypes of S. pneumoniae isolated in this study.

Adolescent↗
PubMed1993

Immune-mediated neuropathies.

Identification of immune-mediated neuropathies is important as these disorders often respond to treatment. Patterns of binding of serum M (monoclonal) proteins and autoantibodies correlate with, and may be diagnostic markers for, specific polyneuropathy syndromes. There is evidence that some of these antibodies may play a pathogenic role in polyneuropathy syndromes. Identification of specific immune polyneuropathy syndromes provides guidance regarding immunosuppressive regimens that may be effective treatments.

Autoantibodies↗