Abstract:
:We developed an efficient electrotransformation system for the pathogenic fungus Histoplasma capsulatum and used it to examine the effects of features of the transforming DNA on transformation efficiency and fate of the transforming DNA and to demonstrate fungal expression of two recombinant Escherichia coli genes, hph and lacZ. Linearized DNA and plasmids containing Histoplasma telomeric sequences showed the greatest transformation efficiencies, while the plasmid vector had no significant effect, nor did the derivation of the selectable URA5 marker (native Histoplasma gene or a heterologous Podospora anserina gene). Electrotransformation resulted in more frequent multimerization, other modification, or possibly chromosomal integration of transforming telomeric plasmids when saturating amounts of DNA were used, but this effect was not observed with smaller amounts of transforming DNA. We developed another selection system using a hygromycin B resistance marker from plasmid pAN7-1, consisting of the E. coli hph gene flanked by Aspergillus nidulans promoter and terminator sequences. Much of the heterologous fungal sequences could be removed without compromising function in H. capsulatum, allowing construction of a substantially smaller effective marker fragment. Transformation efficiency increased when nonselective conditions were maintained for a time after electrotransformation before selection with the protein synthesis inhibitor hygromycin B was imposed. Finally, we constructed a readily detectable and quantifiable reporter gene by fusing Histoplasma URA5 with E. coli lacZ, resulting in expression of functional beta-galactosidase in H. capsulatum. Demonstration of expression of bacterial genes as effective selectable markers and reporters, together with a highly efficient electrotransformation system, provide valuable approaches for molecular genetic analysis and manipulation of H. capsulatum, which have proven useful for examination of targeted gene disruption, regulated gene expression, and potential virulence determinants in this fungus.
journal_name
Infect Immunjournal_title
Infection and immunityauthors
Woods JP,Heinecke EL,Goldman WEdoi
10.1128/IAI.66.4.1697-1707.1998subject
Has Abstractpub_date
1998-04-01 00:00:00pages
1697-707issue
4eissn
0019-9567issn
1098-5522journal_volume
66pub_type
杂志文章abstract::The region of DNA encoding the mannose-fucose-resistant hemagglutinin (MFRHA) of Vibrio cholerae O1 has been localized, and the nucleotide sequence has been determined. The region contains a single open reading frame encoding 230 amino acids, corresponding to a protein of 26.9 kDa. The N terminus of this protein is at...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.61.7.3032-3037.1993
更新日期:1993-07-01 00:00:00
abstract::We examined 205 enterotoxigenic strains of Escherichia coli for colonization factor antigens (CFA) I and II, using an immunodiffusion technique with specific antisera. A total of 36 strains of serogroups O63, O78, O114, O128, and O153 and 1 rough strain possessed CFA/I and gave a single precipitin line; 47 strains of ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.36.1.189-197.1982
更新日期:1982-04-01 00:00:00
abstract::Regulation of the synthesis of Vi polysaccharide, a major virulence determinant in Salmonella enterica serotype Typhi, is under the control of two regulatory systems, ompR-envZ and rscB-rscC, which respond to changes in osmolarity. Some serotype Typhi strains exhibit overexpression of Vi polysaccharide, which masks cl...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00888-06
更新日期:2007-03-01 00:00:00
abstract::Cells of several strains of Streptococcus gordonii attached in much higher numbers to experimental pellicles formed from samples of submandibular or parotid saliva on hydroxyapatite (HA) beads than to buffer controls. The nature of the salivary components responsible were investigated by preparing experimental pellicl...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.59.9.2948-2954.1991
更新日期:1991-09-01 00:00:00
abstract::Tox-+ staphylococcal strains, as opposed to Tox-minus strains, produce epidermal exfoliation within 18 h after direct subcutaneous or intraperitoneal injection into newborn mice. The extracellular product responsible for exfoliation is termed exfoliative toxin (ET). When culture supernatant fluid from the plasmid-cure...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.12.1.193-197.1975
更新日期:1975-07-01 00:00:00
abstract::Mice injected intravenously with a commercially available extract of Coxiella burnetii prepared for use as the antigen in the complement fixation diagnostic test for Q fever were subsequently resistant to infection with Babesia microti, Babesia rodhaini, and Plasmodium vinckei petteri. The parasites appeared to die in...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.24.2.319-325.1979
更新日期:1979-05-01 00:00:00
abstract::An experimental model of coxsackievirus B3 infection in newborn mice was utilized to examine the protective role of antiviral antibodies and an interferon inducer, polyinosinic acid-polycytidylic acid [poly(I:C)]. Subcutaneous administration to the infected mice of specific antiviral antibodies resulted in significant...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.37.2.720-727.1982
更新日期:1982-08-01 00:00:00
abstract::Anti-CD4 antibodies, which cause CD4(+) T-cell depletion, have been shown to increase susceptibility to infections in mice. Thus, development of anti-CD4 antibodies for clinical use raises potential concerns about suppression of host defense mechanisms against pathogens and tumors. The anti-human CD4 antibody kelixima...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.69.2.1032-1043.2001
更新日期:2001-02-01 00:00:00
abstract::The cell surface of Chlamydia psittaci seems important for establishing infection since (i) UV-treated elementary bodies (EB) attach to and are ingested by L cells and (ii) heat or antibody treatment decreases attachment to L cells and promotes the fusion of chlamydiae-containing phagosomes with lysosomes in macrophag...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.40.2.741-751.1983
更新日期:1983-05-01 00:00:00
abstract::Burkholderia cenocepacia strain K56-2 typically has rough colony morphology on agar medium; however, shiny colony variants (shv) can appear spontaneously. These shv all had a minimum of 50% reduction in biomass formation and were generally avirulent in an alfalfa seedling infection model. Three shv-K56-2 S15, K56-2 S7...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00874-07
更新日期:2008-01-01 00:00:00
abstract::Attaching and effacing adhesion to epithelial cells is a pathognomonic feature of infection by both enteropathogenic Escherichia coli (EPEC) and certain strains of verocytotoxin-producing E. coli (VTEC). EPEC adhesion to tissue culture epithelial cells results in activation of the phosphatidylinositol pathway, with el...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.63.9.3316-3326.1995
更新日期:1995-09-01 00:00:00
abstract::Bordetella pertussis, the etiological agent of whooping cough, has the ability to modulate its phenotype in response to environmental conditions by using the BvgAS sensory transduction system which is encoded by the vir locus (now known as bvg). The BvgAS system is part of a large family of two-component sensory trans...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.63.10.3780-3789.1995
更新日期:1995-10-01 00:00:00
abstract::The response to reinfection with Ethiopian Leishmania donovani was evaluated in genetically determined noncure (H-2d) B10.D2 mice that are able to resolve infection due to sublethal irradiation pretreatment after inoculation with a low parasite dose and in C57BL/10 mice that demonstrate the genetically determined cure...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.57.9.2892-2899.1989
更新日期:1989-09-01 00:00:00
abstract::Plasmid DNA from two strains of enterotoxigenic Escherichia coli harboring genes encoding coli surface antigen 4 (CS4) and from seven Indian enterotoxigenic E. coli isolates cross-hybridized at low stringency but not at high stringency with two polynucleotide probes derived from the colonization factor antigen I (CFA/...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.60.9.3799-3806.1992
更新日期:1992-09-01 00:00:00
abstract::In a previous report (Y. Ohmori, S. Hanazawa, S. Amano, T. Miyoshi, K. Hirose, and S. Kitano, infect. Immun. 55:947-954, 1987), we showed that human gingival fibroblasts spontaneously produce thymocyte-activating factor (FTAF), which stimulates mitogen-induced thymocyte proliferation. In the present study, we examined...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.56.1.272-274.1988
更新日期:1988-01-01 00:00:00
abstract::Proliferation of Bordetella pertussis in the lungs of infant mice challenged by the intranasal route was examined. The bacteria rapidly proliferated in the lungs of mice challenged with a sublethal dose of a wild-type strain (BP338) or a filamentous hemagglutinin mutant (BPM409) from 500 at day 0 to 10(7) at day 15. T...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.58.10.3445-3447.1990
更新日期:1990-10-01 00:00:00
abstract::Oral or intravenous inoculation of previously unexposed juvenile and adult ferrets with Campylobacter jejuni uniformly resulted in intestinal colonization lasting 2 to 12 days. Disease varied from mild to moderate diarrhea, which resolved in 2 to 3 days. Orally infected animals developed agglutinin titers of 8 to 256 ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.58.6.1848-1852.1990
更新日期:1990-06-01 00:00:00
abstract::Immune cytolysis was measured by release of absorbed radioactive chromium from infected cells that were incubated with antiviral antibody and complement. The presence of virus-specific antigens detected in this manner on the surface of several types of cells infected with Japanese encephalitis (JE) virus did not corre...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.11.2.330-333.1975
更新日期:1975-02-01 00:00:00
abstract::The human pathogen enterohemorrhagic Escherichia coli (EHEC) O157:H7 has two histidine sensor kinases, QseC and QseE, which respond to the mammalian adrenergic hormones epinephrine and norepinephrine by increasing their autophosphorylation. Although QseC and QseE are present in nonpathogenic strains of E. coli, EHEC e...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.05921-11
更新日期:2012-02-01 00:00:00
abstract::Photobacterium damselae subsp. damselae causes infections and fatal disease in marine animals and in humans. Highly hemolytic strains produce damselysin (Dly) and plasmid-encoded HlyA (HlyA(pl)). These hemolysins are encoded by plasmid pPHDD1 and contribute to hemolysis and virulence for fish and mice. In this study, ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00155-13
更新日期:2013-09-01 00:00:00
abstract::Differences in infection kinetics and host response between Burkholderia multivorans and Burkholderia cenocepacia were demonstrated in a pulmonary infection model in BALB/c mice. B. multivorans persisted in the lung, while B. cenocepacia was cleared. Indirect immunofluorescence and electron microscopy of B. multivoran...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.72.10.6142-6147.2004
更新日期:2004-10-01 00:00:00
abstract::The 120-kDa rickettsial outer membrane protein (rOmpB) is encoded by a gene with the capacity to encode a protein of approximately 168 kDa. The carboxy-terminal end of the molecule is apparently cleaved to yield 120- and 32-kDa products. Both polypeptides are surface exposed and remain associated with the outer membra...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.60.1.159-165.1992
更新日期:1992-01-01 00:00:00
abstract::The opacity (Opa) proteins mediate a variety of interactions between the bacterium Neisseria meningitidis and its human host. These interactions are thought to be of central importance in both the asymptomatic colonization of the nasopharynx and the sporadic occurrence of meningococcal disease. The receptor specificit...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00293-06
更新日期:2006-09-01 00:00:00
abstract::A T antigen preparation free of trypsin was obtained by application of CNBr-Sepharose linked to pure trypsin. Purification on an ion exchange chromatography column results in an electrophoretically homogeneous preparation of T protein. ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.13.3.993-994.1976
更新日期:1976-03-01 00:00:00
abstract::The purpose of our experiment was to evaluate the destructive potential of a strain of Actinomyces viscosus on the periodontium of sensitized rodents, describing the induced lesions on the basis of quantitative cytology. The experimental design comprised in principle the following procedures. Young germfree rats were ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.10.3.565-577.1974
更新日期:1974-09-01 00:00:00
abstract::Salmonella is a widespread zoonotic enteropathogen that causes gastroenteritis and fatal typhoidal disease in mammals. During systemic infection of mice, Salmonella enterica serovar Typhimurium resides and replicates in macrophages within the "Salmonella-containing vacuole" (SCV). It is surprising that the substrates ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00093-09
更新日期:2009-07-01 00:00:00
abstract::Actinobacillus pleuropneumoniae can use porcine transferrin as the sole source of iron. Two proteins with molecular masses of approximately 60 kDa (TfbA) and 110 kDa have been shown to specifically bind porcine transferrin; from the TfbA protein, three isoforms from A. pleuropneumoniae serotypes 1, 5, and 7 have been ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.63.10.3846-3850.1995
更新日期:1995-10-01 00:00:00
abstract::The ultrastructural organization of Mycoplasma pneumoniae membranes and spatial relationships of this pathogen to epithelial cells in tracheal organ cultures were examined ultrastructurally by freeze-fracture techniques. Areas of morphologically distinct cell membrane variability characterized by membrane blebs and al...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:
更新日期:1980-09-01 00:00:00
abstract::The UspA1 and UspA2 proteins of Moraxella catarrhalis are potential vaccine candidates for preventing disease caused by this organism. We have characterized both proteins and evaluated their vaccine potential using both in vitro and in vivo assays. Both proteins were purified from the O35E isolate by Triton X-100 extr...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:
更新日期:1998-09-01 00:00:00
abstract::Mouse fibroblasts (L cells) were infected in suspension with Chlamydia psittaci (6BC) and then plated out on a solid substrate at a density of 80 cells per cm2 so that the effect of chlamydial infection on the division of single host cells and their progeny could be determined. Uninfected L cells multiplied with a mea...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.19.1.281-286.1978
更新日期:1978-01-01 00:00:00