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2. The Yersinia pseudotuberculosis adhesin YadA mediates intimate bacterial attachment to and entry into HEp-2 cells

Author

M C Copass, S Falkow, and James B. Bliska

Subjects
Integrins, Immunology, Integrin, Virulence, Microbiology, Bacterial Adhesion, Plasmid, Bacterial Proteins, Cell–cell interaction, Tumor Cells, Cultured, Humans, Yersinia pseudotuberculosis, Cytotoxic T cell, Adhesins, Bacterial, Cell adhesion, biology, Integrin beta1, biology.organism_classification, Bacterial adhesin, Infectious Diseases, biology.protein, Parasitology, Protein Tyrosine Phosphatases, Bacterial Outer Membrane Proteins, Plasmids, Research Article
Abstract

We characterized a bacterium-host cell interaction that is mediated by the Yersinia adhesin YadA. Derivatives of the virulence plasmid pIB1 harboring mutations in yadA, yopE, or yopH or in a low-calcium-response regulatory locus were introduced into a Yersinia pseudotuberculosis YPIII strain defective for Inv. The mutant strains were tested for the capacity to attach to and enter HEp-2 cells and express the cytotoxic activities of YopE and YopH. As previously shown, expression of YadA was necessary for bacterial attachment and Yop activity in the absence of Inv (R. Rosqvist, A. Forsberg, M. Rimpilainen, T. Bergman, and H. Wolf-Watz, Mol. Microbiol. 4:657-667, 1990). In addition, bacterial entry into HEp-2 cells occurred efficiently when YadA was expressed in the absence of YopE and YopH. These results demonstrated that YadA mediates intimate attachment of Y. pseudotuberculosis to HEp-2 cells and that phagocytic uptake of bacteria by this pathway is inhibited by the synergistic activities of YopH and YopE. A role for beta 1 integrins as host cell receptors for this bacterial attachment and entry mechanism was supported by HEp-2 cell adhesion and monoclonal antibody neutralization studies.

Published
1993

3. Pyrazinamidase, CR-MOX agar, salicin fermentation-esculin hydrolysis, and D-xylose fermentation for identifying pathogenic serotypes of Yersinia enterocolitica

Author

G P Carter, S Falkow, I K Wachsmuth, J. J. Farmer, and Virginia L. Miller

Subjects
Microbiology (medical), Serotype, food.ingredient, biology, Virulence, biology.organism_classification, Enterobacteriaceae, Virology, Microbiology, chemistry.chemical_compound, food, Plasmid, Salicin, chemistry, Agar, Yersinia enterocolitica, Bacteria, Research Article
Abstract

We evaluated several simple laboratory tests that have been used to identify pathogenic serotypes of Yersinia enterocolitica or to indicate the pathogenic potential of individual strains. A total of 100 strains of Y. enterocolitica were studied, including 25 isolated during five outbreak investigations, 63 from sporadic cases, and 12 from stock cultures. The pyrazinamidase test, which does not depend on the Yersinia virulence plasmid, correctly identified 60 of 63 (95% sensitivity) strains of pathogenic serotypes and 34 of 37 (92% specificity) strains of nonpathogenic serotypes. Salicin fermentation-esculin hydrolysis (25 degrees C, 48 h) correctly identified all 63 (100% sensitivity) strains of the pathogenic serotypes and 34 of 37 (92% specificity) strains of the nonpathogenic serotypes. The results of the pyrazinamidase and salicin-esculin tests disagreed for only 7 of the 100 strains of Y. enterocolitica, and these would require additional testing. Congo red-magnesium oxalate (CR-MOX) agar determines Congo red dye uptake and calcium-dependent growth at 36 degrees C, and small red colonies are present only if the strain contains the Yersinia virulence plasmid. This test has proven to be extremely useful for freshly isolated cultures, but only 15 of 62 strains of pathogenic serotypes that had been stored for 1 to 10 years were CR-MOX positive. None of the 16 strains of Y. enterocolitica serotype O3 fermented D-xylose, so this test easily differentiated strains of this serotype, which now appears to be the most common in the United States. Although antisera that can actually be used to serotype strains of Y. enterocolitica are not readily available, the four simple tests described above can be used to screen for pathogenic serotypes.

Published
1992

4. Genetic analysis of Escherichia coli urease genes: evidence for two distinct loci

Author

S Falkow and C. M. Collins

Subjects
DNA, Bacterial, DNA Mutational Analysis, Restriction Mapping, Molecular cloning, medicine.disease_cause, Microbiology, law.invention, Nucleic acid thermodynamics, Plasmid, Restriction map, law, Sequence Homology, Nucleic Acid, Escherichia coli, medicine, Cloning, Molecular, Molecular Biology, biology, Providencia stuartii, Nucleic Acid Hybridization, Gene Expression Regulation, Bacterial, biology.organism_classification, Urease, Molecular biology, Molecular Weight, Blotting, Southern, Restriction enzyme, Genes, Bacterial, DNA Transposable Elements, Recombinant DNA, Research Article, Plasmids
Abstract

Studies with two uropathogenic urease-producing Escherichia coli strains, 1021 and 1440, indicated that the urease genes of each are distinct. Recombinant plasmids encoding urease activity from E. coli 1021 and 1440 differed in their restriction endonuclease cleavage sites and showed minimal DNA hybridization under stringent conditions. The polypeptides encoded by the DNA fragments containing the 1021 and 1440 urease loci differed in electrophoretic mobility under reducing conditions. Regulation of urease gene expression differed in the two ureolytic E. coli. The E. coli 1021 locus is probably chromosomally encoded and has DNA homology to Klebsiella, Citrobacter, Enterobacter, and Serratia species and to about one-half of the urease-producing E. coli tested. The E. coli 1440 locus is plasmid encoded; plasmids with DNA homology to the 1440 locus probe were found in urease-producing Salmonella spp., Providencia stuartii, and two E. coli isolates. In addition, the 1440 urease probe was homologous to Proteus mirabilis DNA.

Published
1990

5. Bacterial epithelial cell cross talk

Author

B, Raupach, J, Mecsas, U, Heczko, S, Falkow, and B B, Finlay

Subjects
Salmonella typhimurium, Colon, Epithelial Cells, Bacterial Adhesion, Yersinia, Peyer's Patches, Enterobacteriaceae, Phagocytosis, Escherichia coli, Animals, Humans, Shigella, Immunity, Mucosal, Bacterial Outer Membrane Proteins
Published
1999

6. Differential trafficking of live and dead Mycobacterium marinum organisms in macrophages

Author

Kathleen George, Lucia P. Barker, P. L. C. Small, and S. Falkow

Subjects
Immunology, Population, Cathepsin D, Vacuole, Microbiology, Cell Line, Mycobacterium, Mice, Phagosomes, Animals, education, Mycobacterium marinum, Phagosome, Latex beads, Cathepsin, education.field_of_study, Microscopy, Confocal, biology, Macrophages, Biological Transport, biology.organism_classification, Cell biology, Infectious Diseases, Parasitology, Research Article
Abstract

We characterized the Mycobacterium marinum phagosome by using a variety of endocytic markers to follow the path of the bacteria through a mouse macrophage cell line. Using a laser confocal microscope, we found that the majority of viable M. marinum cells were in nonacidic vacuoles that did not colocalize with the vacuolar proton ATPase (V-ATPase), the calcium-independent mannose-6-phosphate receptor (CI-M6PR), or cathepsin D. In contrast, heat-killed organisms and latex beads were in acidic vacuoles which contained the V-ATPase, the CI-M6PR, and cathepsin D. A population of vesicles that contained live M. marinum labeled with the lysosomal glycoprotein LAMP-1, but the percentage of vacuoles that labeled was lower than for heat-killed organisms or latex beads. When testing live and heat-killed Mycobacterium tuberculosis, we found levels of colocalization with LAMP- and cathepsin D comparable to those for the M. marinum isolate. We conclude that M. marinum, like M. tuberculosis, can circumvent the host endocytic pathway and reside in an intracellular compartment which is not acidic and does not fuse with lysosomes. In addition, we describe a system for sampling a large population of intracellular organisms by using a laser confocal microscope.

Published
1997

8. Salmonellosis: host immune responses and bacterial virulence determinants

Author

B D, Jones and S, Falkow

Subjects
Salmonella typhimurium, Salmonella Infections, Animal, Virulence, Salmonella Infections, Animals, Humans, Salmonella typhi
Abstract

The lifestyle of bacterial pathogens requires them to establish infection in the face of host immunity. Upon entering a potential host, a variety of interactions are initiated, the outcome of which depends upon a myriad of attributes of each of the participants. In this review we discuss the interactions that occur between pathogenic Salmonella species and the host immune systems, but when appropriate to broaden perspective, we have provided a general overview of the interactions between bacterial pathogens and animal hosts. Pathogenic Salmonella species possess an array of invasion genes that produce proteins secreted by a specialized type III secretion apparatus. These proteins are used by the bacteria to penetrate the intestinal mucosa by invading and destroying specialized epithelial M cells of the Peyer's patches. This maneuver deposits the bacteria directly within the confines of the reticuloendothelial system. The host responds to these actions with nonspecific phagocytic cells and an inflammatory response as well as by activating specific cellular and humoral immune responses. Salmonella responds to this show of force directly. It appears that the bacteria invade and establish a niche within the very cells that have been sent to destroy them. Efforts are underway to characterize the factors that allow these intracellular bacteria to customize intracellular vacuoles for their own purposes. It is the constant play between these interactions that determines the outcome of the host infection, and clearly they will also shape the evolution of new survival strategies for both the bacterium and the host.

Published
1996

10. Isolation of hyperinvasive mutants of Salmonella

Author

C A, Lee and S, Falkow

Subjects
Salmonella typhimurium, Methylnitronitrosoguanidine, Mutagenesis, Insertional, Phenotype, Mutagenesis, DNA Transposable Elements, Tumor Cells, Cultured, Animals, Humans, Salmonella typhi, Cells, Cultured, Epithelium
Published
1994

11. The ail gene of Yersinia enterocolitica has a role in the ability of the organism to survive serum killing

Author

S Falkow and D E Pierson

Subjects
Blood Bactericidal Activity, Immunology, Mutant, Gene Expression, CHO Cells, Yersinia, In Vitro Techniques, Microbiology, Bacterial Adhesion, Transformation, Genetic, Cricetinae, Gene expression, Animals, Humans, RNA, Messenger, Yersinia enterocolitica, Gene, Cells, Cultured, biology, Temperature, biology.organism_classification, Enterobacteriaceae, RNA, Bacterial, Infectious Diseases, Cell culture, Genes, Bacterial, Parasitology, Bacteria, Research Article, Bacterial Outer Membrane Proteins
Abstract

Two Yersinia enterocolitica genes, inv and ail, play a major role in the ability of this microorganism to enter cultured mammalian cells. ail-homologous sequences are present only in pathogenic species and strains of Yersinia. We previously demonstrated (D. E. Pierson and S. Falkow, Infect. Immun. 58:1059-1064, 1990) that four different nonpathogenic isolates of Y. enterocolitica are not able to invade tissue culture cells because they contain functionally inactive variants of the inv gene. When a functional version was introduced into these strains, they became invasive. In this study, we introduced a functional ail gene into the same strains and found that the ail gene was expressed but that these strains neither adhere to nor invade cultured animal cells. However, these recombinant strains became resistant to killing by human serum, whereas their parental strains were not. Using an ail mutant, we also demonstrate that the ail gene has a role in both invasion/adherence and serum resistance in a pathogenic isolate of Y. enterocolitica. These results support a role for Ail in the pathogenesis of Y. enterocolitica infection and disease.

Published
1993

12. Isolation, expression, and nucleotide sequencing of the pilin structural gene of the Brazilian purpuric fever clone of Haemophilus influenzae biogroup aegyptius

Author

J W St Geme rd and S Falkow

Subjects
Haemophilus influenzae biogroup aegyptius, Immunology, Molecular Sequence Data, medicine.disease_cause, Microbiology, Pilus, Haemophilus influenzae, medicine, Humans, Brazilian purpuric fever, Amino Acid Sequence, Cloning, Molecular, Gene, Purpura, Genetics, biology, Base Sequence, Sequence Homology, Amino Acid, Hemagglutination, Structural gene, Nucleic acid sequence, medicine.disease, biology.organism_classification, Blotting, Southern, Infectious Diseases, Oligodeoxyribonucleotides, Genes, Bacterial, Pilin, biology.protein, bacteria, Parasitology, Fimbriae Proteins, Glycoconjugates, Sequence Alignment, Brazil, Research Article, Bacterial Outer Membrane Proteins
Abstract

In this study we isolated the pilin gene from the Brazilian purpuric fever (BPF) clone of Haemophilus influenzae biogroup aegyptius, expressed the gene in Escherichia coli, and determined its nucleotide sequence. Comparison of the nucleotide sequence of the BPF pilin gene with the sequences of pilin genes from strains of H. influenzae sensu stricto demonstrated a high degree of identity. Consistent with this observation, hemagglutination inhibition studies performed with a series of glycoconjugates indicated that BPF pili and H. influenzae type b pili possess the same erythrocyte receptor specificity.

Published
1993

13. How steady are steady-state mountain belts? A reexamination of the Olympic Mountains (Washington state, USA)

Author

L. Michel, C. Glotzbach, S. Falkowski, B. A. Adams, and T. A. Ehlers

Subjects
Dynamic and structural geology, QE500-639.5
Abstract

The Olympic Mountains of Washington state (USA) represent the aerially exposed accretionary wedge of the Cascadia Subduction Zone and are thought to be in flux steady state, whereby the mass outflux (denudation) and influx (tectonic accretion) into the mountain range are balanced. We use a multi-method approach to investigate how temporal variations in the influx and outflux could affect previous interpretations of flux steady state. This includes the analysis of published and new thermochronometric ages for (U–Th) ∕ He dating of apatite and zircon (AHe and ZHe, respectively), fission-track dating of apatite and zircon (AFT and ZFT, respectively), 1-D thermo-kinematic modeling of thermochronometric data, and independent estimates of outflux and influx. In total, we present 61 new AHe, ZHe, AFT, and ZFT thermochronometric ages from 21 new samples. AHe ages are generally young ( 2 to −1 around 5–7 Ma. With the onset of Plio–Pleistocene glaciation, exhumation rates increased to values > 1 km Myr−1. This demonstrates that the material outflux varies through time, requiring a commensurate variation in influx to maintain flux steady state. Evaluation of the offshore and onshore sediment record shows that the material influx is also variable through time and that the amount of accreted sediment in the wedge is spatially variable. This qualitatively suggests that significant perturbations of steady state occur on shorter timescales (105–106 years), like those created by Plio–Pleistocene glaciation. Our quantitative assessment of influx and outflux indicates that the Olympic Mountains could be in flux steady state on long timescales (107 years).

Published
2019
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14. Invasin expression in Yersinia pseudotuberculosis

Author

M Simonet and S Falkow

Subjects
Immunology, Clone (cell biology), Virulence, Spleen, CHO Cells, Biology, Microbiology, Mice, Plasmid, Bacterial Proteins, Cricetinae, medicine, Mesenteric lymph nodes, Yersinia pseudotuberculosis, Animals, Adhesins, Bacterial, Antibodies, Monoclonal, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, Enterobacteriaceae, Infectious Diseases, medicine.anatomical_structure, Parasitology, Bacterial outer membrane, Research Article
Abstract

A 3.2-kb region on the chromosome of Yersinia pseudotuberculosis, called inv, encodes invasin, a 103-kDa protein of the bacterial outer membrane. Invasin mediates bacterial entry into cultured animal cells. Six Y. pseudotuberculosis strains isolated from animal or human infections were analyzed for the presence of inv-related sequences with a radiolabeled inv clone, pRI203. We found that inv-specific sequences were present in all strains studied. Strains cured of virulence plasmid pYV were studied by Western immunoblot analysis with a monoclonal antibody directed against invasin. All but one strain produced invasin, but some strains produced more invasin than others. A strong correlation was found between the level of invasin production by these strains and their ability to enter into HEp-2 or CHO cells. The virulence of these strains was assessed in a murine model by measuring the number of bacteria in the spleen after intravenous challenge or in the mesenteric lymph nodes after intragastric challenge. The capacities of strains to invade cultured mammalian cells and to colonize the spleen were strongly correlative. In contrast, the ability of strains to translocate from the intestinal lumen to the mesenteric lymph nodes after intragastric inoculation did not correlate with their in vitro invasiveness.

Published
1992

15. Identification of uncultured microorganisms: expanding the spectrum of characterized microbial pathogens

Author

D A, Relman and S, Falkow

Subjects
RNA, Bacterial, Bacteria, RNA, Ribosomal, 16S, Animals, Humans, Bacterial Infections, Infections, Microbiology, Polymerase Chain Reaction, Phylogeny
Abstract

The combination of enzymatic nucleic acid amplification techniques with 16S rRNA-based molecular phylogeny has brought about a new approach to the identification of microbial pathogens that can not be cultivated in the laboratory. The applications of this experimental approach to bacillary angiomatosis and to Whipple's disease have revealed the presence of two previously uncharacterized organisms. These results suggest the existence of a far greater microbial diversity among human pathogens than has been so far appreciated with culture-dependent methods. PCR-based studies of aquatic environmental microbial communities have already reached similar conclusions. As a result, new and provocative questions are raised concerning the association of amplified 16S rRNA sequences with diseased tissue. The answers must await the results of further investigations and the expansion of sequence data bases.

Published
1992

17. Characterization of interactions of enteropathogenic Escherichia coli O127:H6 with mammalian cells in vitro

Author

C L, Francis, A E, Jerse, J B, Kaper, and S, Falkow

Subjects
Adhesins, Escherichia coli, Cytochalasin D, Microvilli, Cell Survival, Bacterial Adhesion, Cell Line, Microscopy, Electron, Carcinoma, Squamous Cell, Escherichia coli, Tumor Cells, Cultured, Humans, Colchicine, Laryngeal Neoplasms, Cytoskeleton, Bacterial Outer Membrane Proteins, Plasmids
Abstract

Previous studies have identified two bacterial factors involved in enteropathogenic Escherichia coli (EPEC) infection. A plasmid-mediated EPEC adherence factor (EAF) is responsible for initial and localized adherence. A chromosomally encoded E. coli attachment and effacement factor (eae) is involved in effacement of the eukaryotic cell surface and characteristic "pedestal" formation. By using isogenic strains deficient in either EAF, eae, or both, the process of EPEC adherence and entry in vitro was examined. While EAF proved necessary and sufficient for efficient bacterial association with HEp-2 cells, both EAF and eae were required for efficient effacement of and entry into these cells and other cultured cell lines. Invasion mediated by eae was markedly inhibited by cytochalasin D and colchicine. Afimbrial adhesion or type I pili from uropathogenic strains of E. coli substituted for EAF in EAF-Eae+ strains to provide initial adherence to HEp-2 cells and to facilitate actin condensation.

Published
1991

18. Invasion and replication of Salmonella typhimurium in animal cells

Author

Lorise C. Gahring, B B Finlay, Fred Heffron, and S Falkow

Subjects
Lipopolysaccharides, Salmonella typhimurium, Salmonella, Phagocytosis, Immunology, Mutant, medicine.disease_cause, Microbiology, Bacterial Adhesion, Cell Line, Mice, medicine, Macrophage, Animals, Mutation, biology, Macrophages, biology.organism_classification, Enterobacteriaceae, Epithelium, Infectious Diseases, medicine.anatomical_structure, Cell culture, Parasitology, Research Article
Abstract

A total of 81 avirulent Tn10 insertion mutants of Salmonella typhimurium have previously been described. These mutants were selected for the inability to survive in murine macrophages. We have characterized the abilities of the most avirulent of these mutants to adhere to, invade, and replicate in both macrophages and nonphagocytic epithelial cells. The results suggest that most mutants contain a defect that is specific to survival within professional phagocytes. These mutants invaded and replicated normally within nonphagocytic human colon adenocarcinoma cells (Caco-2) but did not survive in the macrophage cell line J774. One mutant invaded both macrophages and epithelial cells much less efficiently than the parental strain. The defect associated with this mutant appears to be a result of decreased adherence to animal cells.

Published
1990

19. Bordetella pertussis filamentous hemagglutinin: evaluation as a protective antigen and colonization factor in a mouse respiratory infection model

Author

K T Mountzouros, S Falkow, J L Cowell, David A. Relman, and A Kimura

Subjects
Bordetella pertussis, Whooping Cough, Immunology, Filamentous haemagglutinin adhesin, Active immunization, Microbiology, Immunoglobulin G, Mice, medicine, Animals, Lung, Pertussis Vaccine, Antigens, Bacterial, Mice, Inbred BALB C, biology, Immunization, Passive, Respiratory infection, respiratory system, biology.organism_classification, Antibodies, Bacterial, Trachea, Infectious Diseases, medicine.anatomical_structure, Hemagglutinins, Humoral immunity, biology.protein, Pertussis vaccine, Parasitology, Immunization, medicine.drug, Respiratory tract, Research Article
Abstract

Filamentous hemagglutinin (FHA) is a cell surface protein of Bordetella pertussis which functions as an adhesin for this organism. It is a component of many new acellular pertussis vaccines. The proposed role of FHA in immunity to pertussis is based on animal studies which have produced some conflicting results. To clarify this situation, we reexamined the protective activity of FHA in an adult mouse respiratory infection model. Four-week-old BALB/c mice were immunized with one or two doses of 4 or 8 micrograms of FHA and then aerosol challenged with B. pertussis Tohama I. In control mice receiving tetanus toxoid, the CFU in the lungs increased from 10(5) immediately following infection to greater than 10(6) by days 5 and 9 after challenge. Mice immunized with FHA by the intraperitoneal or intramuscular route had significantly reduced bacterial colonization in the lungs. A decrease in colonization of the trachea was also observed in FHA-immunized mice. Evaluation of antibody responses in these mice revealed high titers of immunoglobulin G (IgG) and IgM to FHA in sera and of IgG to FHA in lung lavage fluids. No IgA to FHA was detected. BALB/c mice were also passively immunized intravenously with either goat or rat antibodies to FHA and then aerosol challenged 24 h later, when anti-FHA antibodies were detected in the respiratory tract. Lung and tracheal colonization was markedly reduced in mice immunized with FHA-specific antibodies compared with those receiving control antibodies. In additional studies, the role of FHA in the colonization of the mouse respiratory tract was evaluated by using strain BP101, an FHA mutant of B. pertussis. FHA was important in the initial colonization of the mouse trachea, but was not required for colonization of the trachea later in the infection. FHA was not a factor in colonization of the lungs. Collectively, these experiments demonstrate (i) that systemic immunization with FHA can provide significant protection against B. pertussis infection in both the lower and upper respiratory tract of mice as defined by the lungs and trachea, respectively; (ii) that this protection is mediated primarily by serum antibodies to FHA, which transudate into respiratory secretions; and (iii) that FHA is an important upper respiratory tract colonization factor. These studies provide further evidence for the role of FHA in pertussis pathogenesis and immunity.

Published
1990

20. Risk profiles and one-year outcomes of patients with newly diagnosed atrial fibrillation in India: Insights from the GARFIELD-AF Registry

Author

Jitendra PS. Sawhney, Veerappa A. Kothiwale, Vikas Bisne, Rajashekhar Durgaprasad, Praveen Jadhav, Manoj Chopda, Velam Vanajakshamma, Ramdhan Meena, Govindan Vijayaraghavan, Kamaldeep Chawla, Jagan Allu, Karen S. Pieper, A. John Camm, Ajay K. Kakkar, Jean-Pierre Bassand, David A. Fitzmaurice, Samuel Z. Goldhaber, Shinya Goto, Sylvia Haas, Werner Hacke, Lorenzo G. Mantovani, Frank Misselwitz, Alexander G.G. Turpie, Martin van Eickels, Freek W.A. Verheugt, Gloria Kayani, Keith A.A. Fox, Bernard J. Gersh, Hector Lucas Luciardi, Harry Gibbs, Marianne Brodmann, Frank Cools, Antonio Carlos Pereira Barretto, Stuart J. Connolly, Alex Spyropoulos, John Eikelboom, Ramon Corbalan, Dayi Hu, Petr Jansky, Jørn Dalsgaard Nielsen, Hany Ragy, Pekka Raatikainen, Jean-Yves Le Heuzey, Harald Darius, Matyas Keltai, Sanjay Kakkar, Jitendra Pal Singh Sawhney, Giancarlo Agnelli, Giuseppe Ambrosio, Yukihiro Koretsune, Carlos Jerjes Sánchez Díaz, Hugo Ten Cate, Dan Atar, Janina Stepinska, Elizaveta Panchenko, Toon Wei Lim, Barry Jacobson, Seil Oh, Xavier Viñolas, Marten Rosenqvist, Jan Steffel, Pantep Angchaisuksiri, Ali Oto, Alex Parkhomenko, Wael Al Mahmeed, David Fitzmaurice, D.Y. Hu, K.N. Chen, Y.S. Zhao, H.Q. Zhang, J.Z. Chen, S.P. Cao, D.W. Wang, Y.J. Yang, W.H. Li, Y.H. Yin, G.Z. Tao, P. Yang, Y.M. Chen, S.H. He, Ying Wang, Yong Wang, G.S. Fu, X. Li, T.G. Wu, X.S. Cheng, X.W. Yan, R.P. Zhao, M.S. Chen, L.G. Xiong, P. Chen, Y. Jiao, Y. Guo, L. Xue, F.Z. Wang, H. Li, Z.M. Yang, C.L. Bai, J. Chen, J.Y. Chen, X. Chen, S. Feng, Q.H. Fu, X.J. Gao, W.N. Guo, R.H. He, X.A. He, X.S. Hu, X.F. Huang, B. Li, J. Li, L. Li, Y.H. Li, T.T. Liu, W.L. Liu, Y.Y. Liu, Z.C. Lu, X.L. Luo, T.Y. Ma, J.Q. Peng, X. Sheng, X.J. Shi, Y.H. Sun, G. Tian, K. Wang, L. Wang, R.N. Wu, Q. Xie, R.Y. Xu, J.S. Yang, L.L. Yang, Q. Yang, Y. Ye, H.Y. Yu, J.H. Yu, T. Yu, H. Zhai, Q. Zhan, G.S. Zhang, Q. Zhang, R. Zhang, Y. Zhang, W.Y. Zheng, B. Zhou, Z.H. Zhou, X.Y. Zhu, S. Kakkar, J.P.S. Sawhney, P. Jadhav, R. Durgaprasad, A.G. Ravi Shankar, R.K. Rajput, K. Bhargava, R. Sarma, A. Srinivas, D. Roy, U.M. Nagamalesh, M. Chopda, R. Kishore, G. Kulkarni, P. Chandwani, R.A. Pothiwala, M. Padinhare Purayil, S. Shah, K. Chawla, V.A. Kothiwale, B. Raghuraman, G. Vijayaraghavan, V.M. Vijan, G. Bantwal, V. Bisne, A. Khan, J.B. Gupta, S. Kumar, D. Jain, S. Abraham, D. Adak, A. Barai, H. Begum, P. Bhattacharjee, M. Dargude, D. Davies, B. Deshpande, P. Dhakrao, V. Dhyani, S. Duhan, M. Earath, A. Ganatra, S. Giradkar, V. Jain, R. Karthikeyan, L. Kasala, S. Kaur, S. Krishnappa, A. Lawande, B. Lokesh, N. Madarkar, R. Meena, P. More, D. Naik, K. Prashanth, M. Rao, N.M. Rao, N. Sadhu, D. Shah, M. Sharma, P. Shiva, S. Singhal, S. Suresh, V. Vanajakshamma, S.G. Panse, Y. Koretsune, S. Kanamori, K. Yamamoto, K. Kumagai, Y. Katsuda, K. Sadamatsu, F. Toyota, Y. Mizuno, I. Misumi, H. Noguchi, S. Ando, T. Suetsugu, M. Minamoto, Hiroshi Oda, K. Shiraishi, S. Adachi, K. Chiba, H. Norita, M. Tsuruta, T. Koyanagi, H. Ando, T. Higashi, K. Okada, S. Azakami, S. Komaki, K. Kumeda, T. Murayama, J. Matsumura, Y. Oba, R. Sonoda, K. Goto, K. Minoda, Y. Haraguchi, H. Suefuji, H. Miyagi, H. Kato, Tadashi Nakamura, Tsugihiro Nakamura, H. Nandate, R. Zaitsu, Yoshihisa Fujiura, A. Yoshimura, H. Numata, J. Ogawa, H. Tatematsu, Y. Kamogawa, K. Murakami, Y. Wakasa, M. Yamasawa, H. Maekawa, S. Abe, H. Kihara, S. Tsunoda, Katsumi Saito, Kazuyuki Saito, T. Fudo, K. Obunai, H. Tachibana, I. Oba, T. Kuwahata, S. Higa, M. Gushiken, T. Eto, H. Yoshida, D. Ikeda, Yoshitake Fujiura, M. Ishizawa, M. Nakatsuka, K. Murata, C. Ogurusu, M. Shimoyama, M. Akutsu, I. Takamura, F. Hoshino, N. Yokota, T. Iwao, K. Tsuchida, M. Takeuchi, Y. Hatori, Y. Kitami, Yoichi Nakamura, R. Oyama, M. Ageta, Hiroyuki Oda, Y. Go, K. Mishima, T. Unoki, S. Morii, Yuhei Shiga, H. Sumi, T. Nagatomo, K. Sanno, K. Fujisawa, Y. Atsuchi, T. Nagoshi, T. Seto, T. Tabuchi, M. Kameko, K. Nii, K. Oshiro, H. Takezawa, S. Nagano, N. Miyamoto, M. Iwaki, Yuichiro Nakamura, M. Fujii, M. Okawa, Masahiko Abe, Masatake Abe, Mitsunori Abe, T. Saito, T. Mito, K. Nagao, J. Minami, T. Mita, I. Sakuma, T. Taguchi, S. Marusaki, H. Doi, M. Tanaka, T. Fujito, M. Matsuta, T. Kusumoto, S. Kakinoki, K. Ashida, N. Yoshizawa, J. Agata, O. Arasaki, M. Manita, M. Ikemura, S. Fukuoka, H. Murakami, S. Matsukawa, Y. Hata, T. Taniguchi, T. Ko, H. Kubo, M. Imamaki, M. Akiyama, M. Inagaki, H. Odakura, T. Ueda, Y. Katsube, A. Nakata, H. Watanabe, M. Techigawara, M. Igarashi, K. Taga, T. Kimura, S. Tomimoto, M. Shibuya, M. Nakano, K. Ito, T. Seo, S. Hiramitsu, H. Hosokawa, M. Hoshiai, M. 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Ahmad, W. Willcock, J. Cairns, S. Sathananthan, N. de Kare-Silver, A. Gilliland, E. Strieder, A. Howitt, B. Vishwanathan, N. Bird, D. Gray, M. Clark, J. Bisatt, J. Litchfield, E. Fisher, T. Fooks, A.R. Kelsall, E. Alborough, J. Wakeling, M. Parfitt, K. Milne, S. Rogers, R. Priyadharshan, J.L. Oliver, E. Davies, S. Abushal, M. Jacobs, C. Hutton, N.I. Walls, R. Thompson, C. Chigbo, S.M.A. Zaidi, M. Howard, K.C. Butter, S. Barrow, H. Little, I.U. Haq, L. Gibbons, S. Glencross, A.J. McLeod, K. Poland, C. Mulholland, A. Warke, P. Conn, G. Burns, R.N. Smith, S. Lowe, R. Kamath, H.S. Dau, J. Webster, I. Hodgins, S. Vercoe, P.C. Roome, H. Pinnock, J.R.A. Patel, A. Ali, N. Hart, R. Davies, E. Stuart, C.A. Neden, M. Danielsen, R. Heath, P. Sharma, S. Galloway, C. Hawkins, R. Oliver, M. Aylward, S. Mannion, M. Braddick, D. Edwards, A.C. Rothwell, A. Sabir, F. Choudhary, S. Khalaque, A. Wilson, S. Peters, W. Coulson, N. Roberts, A. Heer, S. Coates, B. Ward, D. Jackson, S. Walton, D. Shepherd, M. Sterry, T. Wong, M. Boon, R. Bunney, R. Haria-Shah, R.T. Baron, S. Davies, T. Schatzberger, N. Hargreaves, T. Stephenson, H. Choi, R. Batson, L. Lucraft, T. Myhill, S. Estifano, D. Geatch, J. Wilkinson, R. Veale, K. Forshaw, T. Davies, K. Zaman, P. Vinson, C. Liley, M. Bandrapalli, P. McGinty, R. Wastling, P. McEleny, A. Beattie, P. Cooke, M. Wong, J. Gunasegaram, M. Pugsley, S. Ahmad, C. A'Court, J. Ayers, J. Bennett, S. Cartwright, S. Dobson, C. Dooldeniya, A. Flynn, R. Fox, J. Goram, A. Halpin, A. Hay, P. Jacobs, L. Jeffers, L. Lomax, I. Munro, R. Muvva, M. Nadaph, K. Powell, S. Randfield, D. Redpath, R. Reed, M. Rickenbach, G. Rogers, P.B. Saunders, C. Seamark, J. Shewring, P. Simmons, H. Simper, H. Stoddart, A. Sword, N. Thomas, A. Thomson, H. Gibbs, A. Blenkhorn, B. Singh, W. Van Gaal, W. Abhayaratna, R. Lehman, P. Roberts-Thomson, J. Kilian, D. Coulshed, A. Catanchin, D. Colquhoun, H. Kiat, D. Eccleston, J. French, L. Zimmett, B. Ayres, T. Phan, P. Blombery, D. Crimmins, D. O'Donnell, A. Choi, P. Astridge, M. Arstall, N. Jepson, M. Binnekamp, A. Lee, J. Rogers, G. Starmer, P. Carroll, J. Faunt, A. Aggarwala, L. Barry, C. Batta, R. Beveridge, A. Black, M. Bonner, J. Boys, E. Buckley, M. Campo, L. Carlton, A. Connelly, B. Conway, D. Cresp, H. Dimitri, S. Dixon, M. Dolman, M. Duroux, M. Eskandari, R. Eslick, A. Ferreira-Jardim, T. Fetahovic, D. Fitzpatrick, R. Geraghty, J. Gibbs, T. Grabek, M.H. Modi, K. Hayes, M.P. Hegde, L. Hesketh, B. Hoffmann, B. Jacobson, K. Johnson, C. Juergens, I. Kassam, V. Lawlor, M. Lehman, S. Lehman, D. Leung, S. Mackay, M. MacKenzie, C. McCarthy, C. McIntosh, L. McKeon, H. Morrison, C. Mussap, J.-D. Myers, V. Nagalingam, G. Oldfield, V. O'May, J. Palmer, L. Parsons, K. Patching, T. Patching, V. Paul, M. Plotz, S. Preston, H. Rashad, M. Ratcliffe, S. Raynes, J. Rose, L. Sanders, M. Seremetkoska, H. Setio, S. Shone, P. Shrestha, C. Singh, C. Singleton, N. Stoyanov, S. Sutcliffe, K. Swaraj, J. Tarrant, S. Thompson, I.M. Tsay, M. Vorster, A. Waldman, L. Wallis, E. Wilford, K. Wong, S.J. Connolly, A. Spyropoulos, J. Eikelboom, R. Luton, M. Gupta, A.S. Pandey, S. Cheung, R. Leader, P. Beaudry, F. Ayala-Paredes, J. Berlingieri, J. Heath, G. Poirier, M. Du Preez, R. Nadeau, G. Dresser, R. Dhillon, T. Hruczkowski, B. Schweitzer, B. Coutu, P. Angaran, P. MacDonald, S. Vizel, S. Fikry, R. Parkash, A. Lavoie, J. Cha, B. Ramjattan, J. Bonet, K. Ahmad, L. Aro, T. Aves, K. Beaudry, C. Bergeron, J. Bigcanoe, N. Bignell, L. Breakwell, E. Burke, L. Carroll, B. Clarke, T. Cleveland, S. Daheb, P. Dehghani, I. Denis, Z. Djaidani, P. Dorian, S. Douglass, J. Dunnigan, A. Ewert, D. Farquhar, A. Fearon, L. Ferleyko, D. Fournier, B. Fox, M.-C. Grenier, W. Gulliver, K. Haveman, C. Hines, K. Hines, A.M. Jackson, C. Jean, G. Jethoo, R. Kahlon, S. Kelly, R. Kim, V. Korley, J. Kornder, L. Kwan, J. Largy, C. Lewis, S. Lewis, I. Mangat, R. Moor, J. Navratil, I. Neas, J. Otis, R. Otis, M. Pandey, F. Petrie, A. Pinter, M. Raines, P. Roberts, M. Robinson, G. Sas, S. Schulman, L. Snell, S. Spearson, J. Stevenson, T. Trahey, S. Wong, D. Wright, H. Ragy, A. Abd El-Aziz, S.K. Abou Seif, M.G. El Din, S. El Etriby, A. Elbahry, A. El-Etreby, M. Elkhadem, A. Katta, T. Khairy, A. Mowafy, M. Nawar, A. Ohanissian, A. Reda, M. Reda, H. Salem, N. Sami, S. Samir, M. Setiha, M. Sobhy, A. Soliman, N. Taha, M. Tawfik, E. Zaatout, D. Kettles, J. Bayat, H. Siebert, A. Horak, Y. Kelfkens, R. Garda, T. Pillay, M. Guerra, L. van Zyl, H. Theron, A. Murray, R. Louw, D. Greyling, P. Mntla, V. Ueckermann, R. Loghdey, S. Ismail, F. Ahmed, J. Engelbrecht, A. Ramdass, S. Maharajh, W. Oosthuysen, G. Angel, C. Bester, M. Booysen, C. Boshoff, C. Cannon, S. Cassimjee, C. Chami, G. Conway, A. Davids, L. de Meyer, G. Du Plessis, T. Ellis, L. Henley, M. Karsten, E. Loyd, J. Marks, L. Mavhusa, M. Mostert, A. Page, L. Rikhotso, M. Salie, J. Sasto, F. Shaik, A. Skein, L. Smith, G. Tarr, T. Tau, F. van Zyl, W. Al Mahmeed, G. Yousef, A. Agrawal, M. Nathani, M. Ibrahim, E.M. Esheiba, R. Singh, A. Naguib, M. Abu-Mahfouz, M. Al Omairi, A. Al Naeemi, R. Maruthanayagam, N. Bazargani, A. Wassef, R. Gupta, M. Khan, B. Subbaraman, A. Abdul, A. Al Mulla, S. El Bardisy, P. Haridas, S. Jadhav, K. Magdaluyo, M. Makdad, I. Maqsood, R. Mohamed, N. Sharma, R. Sharma, M. Thanzeel, S.Z. Goldhaber, R. Canosa, P. Rama, E. Blumberg, J. Garcia, P. Mullen, V. Wilson, A. Quick, K. Ferrick, W.M. Kutayli, M. Cox, M. Franco, S. Falkowski, R. Mendelson, M. Williams, S. Miller, S. Beach, A. Alfieri, T. Gutowski, I. Haque, R. Reddy, W. Ahmed, P. Delafontaine, D. Diercks, D. Theodoro, K. Remmel, M. Alberts, R. Ison, H. Noveck, P. Duffy, S. Pitta, D. Nishijima, C. Treasure, N. Asafu-Adjaye, K. Ball, M. Bartlett, M. Bentley, S. Bowers, A. Brown, A. Browne, J. Cameron-Watts, M. Canova, D. Cassidy, K. Cervellione, S. Congal, J. DePauw, A. Dickerson, M. Eley, L. Evans, S. Felpel, K. Ferdinand, D. Fielder, P. Gentry, A. Haideri, F. Hakimi, T. Harbour, E. Hartranft, B. Hawkins, M. Headlee, L. Henson, C. Herrick, T. Hicks, S. Jasinski, A. Jones, L. Jones, P. Jones, S. Karl, M. Keeling, J. Kerr, P. Knowles, J. Langdon, M. Lay, J.A. Lee, T. Lincoln, E. Malone, A. Merliss, D. Merritt, J. Minardo, B. Mooso, C. Orosco, V. Palumbo, M. Parker, T. Parrott, S. Paserchia, G. Pearl, J. Peterson, N. Pickelsimer, T. Purcell, J. Raynor, S. Raziano, C. Richard, T. Richardson, C. Robertson, A. Sage, T. Sanghera, P. Shaw, J. Shoemaker, K. Smith, B. Stephanie, A. Thatcher, H. Theobald, N. Thompson, L. Treasure, T. Tripti, C. Verdi, and V. Worthy

Subjects
Surgery, RD1-811, Diseases of the circulatory (Cardiovascular) system, RC666-701
Abstract

Background: The Global Anticoagulant Registry in the FIELD–Atrial Fibrillation (GARFIELD-AF) is an ongoing prospective noninterventional registry, which is providing important information on the baseline characteristics, treatment patterns, and 1-year outcomes in patients with newly diagnosed non-valvular atrial fibrillation (NVAF). This report describes data from Indian patients recruited in this registry. Methods and results: A total of 52,014 patients with newly diagnosed AF were enrolled globally; of these, 1388 patients were recruited from 26 sites within India (2012–2016). In India, the mean age was 65.8 years at diagnosis of NVAF. Hypertension was the most prevalent risk factor for AF, present in 68.5% of patients from India and in 76.3% of patients globally (P

Published
2018
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21. Horizontal insulating barriers as a way to protect groundwater

Author

R. Cicha-Szot, K. Labus, S. Falkowicz, and N. Madetko

Subjects
Environmental sciences, GE1-350, Geology, QE1-996.5
Abstract

Trenchless Technology of Forming Horizontal Insulating Barriers (TFHB) can be considered a method of groundwater protection against inflow of pollutants. In TFHB technology, the working fluid (sodium silicate solution) and the gelling agent (CO2) are injected separately, using one tool, to different zones of the aquifer profile. Carbon dioxide injected into the saturation zone rises due to buoyancy forces and reaches the silicate which was injected at the water table level. This initiates the process of silicate gelation, resulting in the formation of an insulating barrier. For technological purposes, the gelation time must be controlled, and the resulting gel must have certain mechanical properties. In order to apply THFB in real conditions it was necessary to identify important technological and technical parameters, as well as to define interactions between the injected fluid and the aquifer rocks. Geochemical modelling (equilibrium, reaction path and reactive transport) was used to identify potential geochemical effects of the application of TFHB in sandy aquifers. Certain petrophysical parameters and mineralogical assemblages of aquifers were addressed, taking into account both low and strongly mineralized groundwater. The simulations revealed that TFHB does not have a negative impact on the chemistry of rock-water systems described in this work.

Published
2018
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22. Reply

Author

D. Monack and S. Falkow

Subjects
Infectious Diseases, Immunology and Allergy
Published
1990

23. Nucleotide sequence of pilA, the gene encoding the structural component of type 1 pili in Escherichia coli

Author

S Falkow and P E Orndorff

Subjects
Genetics, Base Sequence, biology, Base pair, Nucleic acid sequence, Protein primary structure, medicine.disease_cause, Microbiology, Pilus, Biochemistry, Genes, Bacterial, Fimbriae, Bacterial, Pilin, Escherichia coli, medicine, biology.protein, Amino Acid Sequence, Fimbriae Proteins, Molecular Biology, Peptide sequence, Gene, Bacterial Outer Membrane Proteins, Research Article
Abstract

The pilA gene of Escherichia coli J96 encoding pilin, the structural component of type 1 pili, was sequenced and found to specify a polypeptide 159 amino acids long preceded by a 23-amino-acid signal peptide. As determined from the DNA sequence, the mature peptide lacked tryptophane and methionine, two amino acids previously shown to be lacking in type 1 pili from E. coli. Also, the amino-terminal sequence of amino acids inferred from the DNA sequence corresponded to earlier 20-amino-acid amino-terminal sequences determined by protein sequencing. In addition, piliation was abolished after a mutation was introduced into the pilA coding region in vitro. A possible site for initiation of transcription and a possible site encoding translation initiation were suggested 85 and 7 base pairs, respectively, from the pilA start codon. There appeared to be scant DNA sequence homology and scant amino acid sequence homology between type 1 pilin and other pilin species isolated from uropathogenic and enteropathogenic E. coli.

Published
1985

24. DNA Hybridization Technique for the Detection of Neisseria gonorrhoeae in Men with Urethritis

Author

King K. Holmes, J S Knapp, Patricia A. Totten, Peter L. Perine, H. Hunter Handsfield, and S. Falkow

Subjects
DNA, Bacterial, Male, Biology, medicine.disease_cause, Microbiology, Gonorrhea, Nucleic acid thermodynamics, chemistry.chemical_compound, Plasmid, Vancomycin, medicine, Citrulline, Humans, Immunology and Allergy, Urethritis, DNA–DNA hybridization, Nucleic Acid Hybridization, Uracil, medicine.disease, Virology, Neisseria gonorrhoeae, Infectious Diseases, chemistry, Phenazines, Gentian Violet, DNA, Plasmids
Abstract

A technique to detect Neisseria gonorrhoeae directly in clinical specimens was developed using a modified DNA-hybridization method. It uses the gonococcal cryptic plasmid as the radiolabeled probe, can detect as few as 100 colony-forming units of N gonorrhoeae or as little as 0.1 pg of purified gonococcal plasmid DNA, and is highly specific. This technique for differentiating between gonococcal and nongonococcal urethritis was evaluated in men with symptomatic urethritis in Seattle. Sixty-three (89%) of 71 who had cultures positive for N gonorrhoeae were also positive by DNA hybridization, and all 42 whose cultures were negative were also negative by DNA hybridization. Five of six isolates from patients who were positive by culture but negative by hybridization lacked the gonococcal cryptic plasmid and belonged to a unique auxo-type which requires proline, citrulline, and uracil for growth.

Published
1983

25. Evaluation of DNA colony hybridization and other techniques for detection of virulence in Yersinia species

Author

Roy M. Robins-Browne, M D Miliotis, Virginia L. Miller, S. Cianciosi, J G Morris, and S Falkow

Subjects
DNA, Bacterial, Microbiology (medical), Restriction Mapping, Virulence, Yersinia, Amidohydrolases, Microbiology, Plasmid, Species Specificity, Animals, Humans, Cloning, Molecular, Yersinia enterocolitica, Genetics, biology, Hybridization probe, Nucleic Acid Hybridization, biology.organism_classification, Enterobacteriaceae, Blotting, Southern, bacteria, Calcium, Gentian Violet, DNA Probes, Molecular probe, Bacteria, Plasmids, Research Article
Abstract

The virulence of yersiniae varies according to (i) species and biotype and (ii) possession of a 67- to 72-kilobase virulence plasmid. Y. pestis, Y. pseudotuberculosis, and biotypes 1B, 2, 3, 4, and 5 of Y. enterocolitica are inherently virulent but express full virulence only when in possession of a virulence plasmid. Other Yersinia species and biotypes 1A and 3B of Y. enterocolitica are seldom implicated in disease. In this study, we prepared DNA probes from eight nonoverlapping regions of the virulence plasmid of a strain of Y. enterocolitica and from the inv and ail chromosomal loci responsible for the invasive capacity of Y. enterocolitica and Y. pseudotuberculosis. The probes were used in colony hybridization experiments to investigate 156 yersiniae of various species and biotypes and of differing virulence. Probes prepared from the inv gene of Y. pseudotuberculosis hybridized with Y. pseudotuberculosis and Y. pestis only, whereas an analogous probe prepared from Y. enterocolitica hybridized with all species and biotypes of yersiniae (but not with other bacteria) regardless of virulence or potential virulence. Probes prepared from the ail region of Y. enterocolitica reacted almost exclusively with Y. enterocolitica strains of pathogenic biotypes. Probes prepared from the virulence plasmid of a serogroup O:8, biotype 1B isolate of Y. enterocolitica identified virulent yersiniae in all species with a high degree of sensitivity and specificity. These probes did not react with yersiniae of avirulent biotypes or species. Of the other assays of virulence evaluated (calcium dependence, binding of crystal violet, and pyrazinamidase activity), binding of crystal violet provided a simple means for identifying plasmid-bearing strains.

Published
1989

26. Genetic Analysis of Essential Plasmid Determinants of Pathogenicity in Yersinia pestis

Author

H. F. Blank, D. A. Portnoy, D. T. Kingsbury, and S. Falkow

Subjects
Transposable element, Yersinia pestis, Mutant, Virulence, medicine.disease_cause, Microbiology, Mice, Plasmid, medicine, Animals, Immunology and Allergy, Escherichia coli, Gene, Antigens, Bacterial, biology, DNA Restriction Enzymes, biology.organism_classification, Molecular biology, Restriction enzyme, Infectious Diseases, Mutation, DNA Transposable Elements, bacteria, Calcium, Plasmids
Abstract

The role of the Yersinia pestis virulence-associated plasmid, pYV019, in the expression of Ca++ dependence, virulence, and the production of the V antigen was investigated. Derivatives of bacteriophage P1 were used to deliver the transposon Tn5 into Y pestis strain EV76. Ca++-independent mutants in which transposon Tn5 had been inserted into plasmid pYV019 were isolated, the resulting plasmids--pYV019::Tn5--were transformed into an Escherichia coli K12 derivative, and the site of insertion of transposon Tn5 was mapped with restriction endonucleases. The plasmids were then transduced by bacteriophage P1 into avirulent strain 195-P1 of Y pestis. The transductants were analyzed for expression of Ca++ dependence, virulence in mice, and the expression of the V antigen. Introduction of plasmid pYV019 with insertions outside of the Ca++ dependence loci restored strain 195-P1 to full virulence, while disruption of plasmid genes associated with Ca++ dependence led to loss of virulence. Using Western blotting analysis and E coli minicells, it was shown that genes specific for the V antigen are plasmid encoded.

Published
1983

27. Cloning of the recA gene of Neisseria gonorrhoeae and construction of gonococcal recA mutants

Author

S Falkow and J M Koomey

Subjects
Transposable element, Genotype, Ultraviolet Rays, DNA repair, Mutant, Mutagenesis (molecular biology technique), Biology, medicine.disease_cause, Microbiology, Plasmid, medicine, Cloning, Molecular, Molecular Biology, Escherichia coli, Genetics, biochemical phenomena, metabolism, and nutrition, Molecular biology, Neisseria gonorrhoeae, Complementation, Rec A Recombinases, Phenotype, Genes, Genes, Bacterial, Mutation, bacteria, Homologous recombination, Research Article, Plasmids
Abstract

Interspecific complementation of an Escherichia coli recA mutant was used to identify recombinant plasmids within a genomic cosmid library derived from Neisseria gonorrhoeae that carry the gonococcal recA gene. These plasmids complement the E. coli recA mutation in both homologous recombination functions and resistance to DNA damaging agents. Subcloning, deletion mapping, and transposon Tn5 mutagenesis were used to localize the gonococcal gene responsible for suppression of the E. coli RecA- phenotype. Defined mutations in and near the cloned gonococcal recA gene were constructed in vitro and concurrently associated with a selectable genetic marker for N. gonorrhoeae and the mutated alleles were then reintroduced into the gonococcal chromosome by transformation-mediated marker rescue. This work resulted in the construction of two isogenic strains of N. gonorrhoeae, one of which expresses a reduced proficiency in homologous recombination activity and DNA repair function while the other displays an absolute deficiency in these capacities. These gonococcal mutants behaved similarly to recA mutants of other procaryotic species and displayed phenotypes consistent with the data obtained by heterospecific complementation in an E. coli recA host. The functional activities of the recA gene products of N. gonorrhoeae and E. coli appear to be highly conserved.

Published
1987

28. Organization and expression of genes responsible for type 1 piliation in Escherichia coli

Author

P E Orndorff and S Falkow

Subjects
Pilus assembly, DNA, Recombinant, Biology, medicine.disease_cause, Microbiology, Pilus, Insertional mutagenesis, Plasmid, Escherichia coli, medicine, Cloning, Molecular, Molecular Biology, Gene, Genetic transfer, DNA Restriction Enzymes, Molecular biology, Phenotype, Genes, Bacterial, Fimbriae, Bacterial, Pilin, Mutation, biology.protein, bacteria, Chromosome Deletion, Plasmids, Research Article
Abstract

The genetic organization of a segment of recombinant DNA conferring the capacity of synthesize E. coli type 1 pili was examined. This 11.2-kilobase (kb) segment of DNA, derived from a clinical isolate, conferred a piliated phenotype (Pil+) on a nonpiliated (Pil-) strain of E. coli K-12 that lacked DNA homologous to the 11.2-kb region. Insertional mutagenesis, deletion mutagenesis, and subcloning of various regions of the 11.2-kb fragment allowed the localization of five genes, each encoding a polypeptide, that were associated with pilus expression. Three gene products, 17, 86, and 30 kilodaltons (kd) in size, were involved in pilus assembly; assembly of the 17-kd structural (pilin) protein into pili was not seen in mutants lacking either the 86- or 30-kd proteins, but pilin synthesis and proteolytic processing were not affected. The fourth polypeptide, 23 kd in size, appeared to be involved in the regulation of pilus expression because mutants lacking this protein exhibited a 40-fold increase in the amount of pilin antigen per cell. The last protein, 14 kd in size, was not associated with piliation by genetic criteria; however, the 14-kd protein was immunoprecipitated with pili, suggesting an association with pili or immunological cross-reactivity with pilin. Immunoprecipitates of minicell transcription translation products revealed that pilus polymerization was taking place in minicells. This may facilitate the study of the molecular steps in pilus biosynthesis and, as a consequence, provide clues to the assembly of supramolecular structures in general.

Published
1984

29. Molecular epidemiology of Vibrio cholerae in the U.S. Gulf Coast

Author

S Falkow, N C Roberts, H B Bradford, and J B Kaper

Subjects
DNA, Bacterial, Microbiology (medical), Molecular epidemiology, Cholera toxin, Nucleic Acid Hybridization, Biology, Louisiana, medicine.disease_cause, medicine.disease, biology.organism_classification, Cholera, Virology, El Tor, Microbiology, Restriction enzyme, Vibrio cholerae, medicine, Humans, Escherichia coli, Research Article, Southern blot
Abstract

Enterotoxigenic strains of Vibrio cholerae O-1, biotype El Tor, isolated from a case of cholera in Texas in 1973, an outbreak of cholera in Louisiana in 1978, and Louisiana sewage samples in 1980 and 1981 were analyzed for their genetic similarities. Chromosomal DNA was isolated from each strain, digested with restriction endonuclease, and analyzed by the Southern blot technique. A radioactive probe consisting of Escherichia coli heat-labile enterotoxin DNA detected cholera toxin gene sequences in these strains and demonstrated that the toxin gene sequence, if not the entire chromosomal DNA, is identical in these strains and distinctly different from other strains of V. cholerae isolated throughout the world. In addition, two strains of enterotoxigenic V. cholerae non-O-1 isolated from clinical cases, were analyzed and found to possess cholera toxin genes which differed in the DNA sequence from the V. cholerae O-1 strains. We concluded that a single strain of enterotoxigenic V. cholerae O-1 is resident in the U.S. Gulf Coast and that a second reservoir of cholera toxin genes exists in V. cholerae non-O-1 strains in Louisiana.

Published
1982

30. Plasmid-mediated properties of a heat-stable enterotoxin-producing Escherichia coli associated with infantile diarrhea

Author

R W Ryder, S Falkow, and I K Wachsmuth

Subjects
Hot Temperature, medicine.drug_class, Immunology, Antibiotics, Extrachromosomal Inheritance, Microbial Sensitivity Tests, Enterotoxin, Biology, medicine.disease_cause, Microbiology, Enterotoxins, Mice, Antibiotic resistance, Plasmid, medicine, Animals, Humans, Heat-stable enterotoxin, Infantile diarrhea, Escherichia coli, Escherichia coli Infections, Infant, Virology, Infectious Diseases, Conjugation, Genetic, Diarrhea, Infantile, Parasitology, Rabbits, Research Article, Plasmids
Abstract

The plasmid mediation and transmissibility of heat-stable enterotoxin production and multiple antibiotic resistance have been demonstrated for Escherichia coli O78:K80:H12 epidemiologically incriminated in a hospital outbreak of infantile diarrhea. The conjugal transfer of a 67 X 10(6) - and a 30 X 10(6)-dalton plasmid was associated with the transfer of resistances and enterotoxin production, respectively. Using antibiotics to select E. coli K-12 transconjugants from a one-step bacterial cross, all of the monitored resistances were transferred concurrently, and 36% of the resistant transconjugants produced enterotoxin.

Published
1976

31. Characterization of plasmids that encode for the K88 colonization antigen

Author

P L Shipley, C L Gyles, and S Falkow

Subjects
Diarrhea, Genetic Linkage, Swine, Immunology, Biology, medicine.disease_cause, Microbiology, Virulence factor, chemistry.chemical_compound, Raffinose, Plasmid, Escherichia coli, medicine, Animals, Enteropathogenic Escherichia coli, Antigen Gene, Gene, Swine Diseases, Antigens, Bacterial, Base Sequence, Molecular biology, Infectious Diseases, Genes, chemistry, Parasitology, DNA, Plasmids, Research Article
Abstract

K88 antigen, and important virulence factor in porcine enteropathogenic Escherichia coli (EEC), can be transferred along with the ability to ferment the trisaccharide raffinose (Raf). The plasmids from a number of EEC strains that encode these two properties were isolated and characterized. In most strains the K88 and Raf genes were found on a single nonconjugative plasmid approximately 50 x 10(6) daltons in size. This plasmid core was conserved with only slight variation among the strains tested. In some transconjugants, larger conjugative plasmids were observed that were apparently recombinants between the Raf/K88 plasmid and a transfer fa(tor. Occasionally plasmids carrying only the raffinose fermentation genes arose by deletion of a deoxyribonucleic acid segment of about 20 x 10(6) daltons that included the K88 antigen gene(s).

Published
1978

32. Cistrons encoding Escherichia coli heat-labile toxin

Author

D M Gill, W S Dallas, and S Falkow

Subjects
DNA, Bacterial, Erythrocytes, Transcription, Genetic, Bacterial Toxins, DNA, Recombinant, Cloning vector, Biology, medicine.disease_cause, Microbiology, DNA sequencing, law.invention, chemistry.chemical_compound, Plasmid, law, Escherichia coli, medicine, Animals, Columbidae, Molecular Biology, Gene, DNA Restriction Enzymes, Molecular biology, Enzyme Activation, Molecular Weight, Genes, chemistry, Protein Biosynthesis, Recombinant DNA, DNA, In vitro recombination, Research Article, Adenylyl Cyclases, Plasmids
Abstract

The structure and products of the two cistrons encoding the Escherichia coli heat-labile toxin (LT) were studied. The LT deoxyribonucleic acid (DNA) region had been isolated as part of a DNA fragment from the plasmid P307, and this fragment was joined to the cloning vector pBR313. Deletion mutations of various lengths were introduced into the LT DNA region and into the adjacent DNA sequences. Analysis of the deletions indicated that the maximum size of the LT DNA region was 1.2 x 10(6) daltons. Two proteins of 11,500 daltons and 25,500 daltons had been shown to be encoded by the LT DNA region. The functions of these LT gene products were investigated. The 11,500-dalton protein had an adsorption activity for Y-1 adrenal cells, and this protein was shown to form aggregates of four or five monomers. The 25,500-dalton protein was shown to have an adenylate cyclase-activating activity. The two cistrons encoding for each of the LT proteins have been located on a genetic map of the LT DNA region. Both cistrons are probably transcribed from the same promoter.

Published
1979

33. Development of a tissue culture model for gonococcal invasion

Author

G. F. Brooks, J. H. Shaw, F. Hayes, and S. Falkow

Subjects
Lysis, Adenocarcinoma, Matrix (biology), urologic and male genital diseases, medicine.disease_cause, Microbiology, Bacterial Adhesion, Tissue culture, stomatognathic system, Tumor Cells, Cultured, medicine, Humans, Molecular Biology, Microvilli, biology, Neisseria meningitidis, General Medicine, biology.organism_classification, Neisseria gonorrhoeae, Mitochondria, Microscopy, Electron, Cytoplasm, Cellular Debris, Bacteria
Abstract

Neisseria gonorrhoeae invasion of the human endometrial cell line HecIB was monitored by electron microscopy. Within six hours postinfection, the gonococci have attached to the surface of some HecIB cells and are embraced by microvilli. Gonococci subsequently enter the HecIB cells in membrane bound vesicles but by eight hours, gonococci can be seen free in the cytoplasm. At twelve hours post-infection some HecIB cells are observed containing hundreds of internalized bacteria. At twenty-four hours gonococci appear in large clusters embedded in a matrix of cellular debris, which are possibly the remains of lysed infected cells. In contrast, N. lactamica is adherent to the monolayer but noninvasive.

Published
1987

34. Polynucleotide sequence relationships among Ent plasmids and the relationship between Ent and other plasmids

Author

J H Crosa, S Falkow, and Magdalene So

Subjects
DNA, Bacterial, Guanine, Hot Temperature, Swine, Polynucleotides, Extrachromosomal Inheritance, Biology, Tritium, medicine.disease_cause, Microbiology, Cytosine, Enterotoxins, chemistry.chemical_compound, Nucleic acid thermodynamics, Plasmid, Escherichia coli, medicine, Animals, Humans, Molecular Biology, Genetics, Mutation, Base Sequence, Nucleic Acid Hybridization, Drug Resistance, Microbial, Molecular biology, Anti-Bacterial Agents, Thymine, Molecular Weight, chemistry, Polynucleotide, Research Article
Abstract

Deoxyribonucleic acid-deoxyribonucleic acid hybridization studies reveal that the plasmids coding for the production of heat stable and heat labile enteroxtoxins of Escherichia coli, regardless of their origin, have a majority of their polynucleotide sequences in common, but are not related in any significant way to those plasmids coding for the synthesis of only ST toxin. The heat stable and heat labile plasmids also share a significant degree of their polynucleotide sequences with plasmids of the FI and FII incompatibility groups, but not with R factors belonging to the I, N, W, P, or X incompatibility groups.

Published
1975

35. Characterization of ampicillin resistance plasmids from Haemophilus ducreyi

Author

S. Falkow, King K. Holmes, H. Hunter Handsfield, Patricia A. Totten, and D. Peters

Subjects
DNA, Bacterial, Transposable element, Penicillin Resistance, R Factors, Biology, medicine.disease_cause, Microbiology, Haemophilus ducreyi, Nucleic acid thermodynamics, Plasmid, Amp resistance, Ampicillin, medicine, Pharmacology (medical), Escherichia coli, Pharmacology, Genetics, Nucleic Acid Hybridization, DNA Restriction Enzymes, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, Restriction enzyme, Infectious Diseases, Autoradiography, bacteria, Transformation, Bacterial, Research Article, medicine.drug
Abstract

Seven strains of Haemophilus ducreyi from diverse geographic origins were analyzed for their plasmid content. All strains were multiply resistant, but only resistance to ampicillin was transferred to Escherichia coli by transformation. The H. ducreyi plasmids encoding for ampicillin resistance were 7.4, 5.7, and 3.6 megadaltons and encoded for part or all of TnA, and ampicillin transposon. The relatedness of these plasmids was examined by restriction endonuclease digestion and DNA-DNA homology with isolated DNA fragments from TnA.

Published
1982

36. Patents and literature

Author

Robert J. Linhardt, S. Falkow, S. L. Moseley, D. Gillespie, I. Brodsky, J. Bresser, J. R. Kiovsky, C. L. Hendrick, S. Lavi, P. Leder, R. Maas, R. A. Owens, T. O. Diener, D. F. Rippe, H. Rubin, D. A. Shafritz, G. M. Wahl, G. R. Stark, R. A. Weinberg, C. J. Tobin, S. M. Bradley, L. B. Wilson, J. T. Wilson, and R. F. Geever

Subjects
business.industry, Hybridization probe, Bioengineering, General Medicine, Computational biology, Biology, business, Molecular Biology, Applied Microbiology and Biotechnology, Biochemistry, Biotechnology
Published
1986

37. Method for the genetic labeling of cryptic plasmids

Author

S Falkow, Fred Heffron, and Magdalene So

Subjects
Genetics, F-factor, Biology, medicine.disease_cause, Microbiology, Phenotype, Transposition (music), F Factor, Plasmid, Conjugation, Genetic, Escherichia coli, otorhinolaryngologic diseases, medicine, Molecular Biology, Plasmids, Research Article
Abstract

A recently developed method for detecting transposition was employed to genetically "label" conjugative plasmids such as F and Ent P307, which do not normally exhibit a readily identifiable phenotype.

Published
1978

38. Analysis of expression and thermoregulation of the Yersinia pseudotuberculosis inv gene with hybrid proteins

Author

Ralph R. Isberg, S Falkow, and A Swain

Subjects
Recombinant Fusion Proteins, Two-hybrid screening, DNA Mutational Analysis, Immunology, lac operon, Locus (genetics), Microbiology, Fusion gene, Bacterial Proteins, Gene expression, Escherichia coli, Yersinia pseudotuberculosis, Cloning, Molecular, Adhesins, Bacterial, Gene, Genetics, Regulation of gene expression, biology, Temperature, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, Molecular biology, Molecular Weight, Infectious Diseases, Gene Expression Regulation, Protein Biosynthesis, DNA Transposable Elements, Parasitology, Research Article
Abstract

A series of translational fusions between the Yersinia pseudotuberculosis inv locus and lacZ was constructed. Each Lac+ fusion strain expressed a hybrid protein containing invasin, the product of the inv locus, at its amino-terminal end. Analysis of these gene fusions allowed determination of the direction of translation of the inv gene. Previous studies of Y. pseudotuberculosis invasion have shown that entry into animal cells is temperature regulated. It is shown here that control of expression of the inv gene is also temperature regulated. phoA gene fusions to inv, when present in Y. pseudotuberculosis, were expressed at lower levels when bacteria were grown at 37 degrees C rather than at 28 degrees C. Similar fusions, in contrast, were regulated in a temperature-independent fashion in Escherichia coli, as was the wild-type inv gene. This implies that Y. pseudotuberculosis has chromosomally encoded trans-acting functions that normally thermoregulate expression of inv.

Published
1988

40. Location of the piliation factor on the chromosome of Escherichia coli

Author

S. Falkow, L.S. Baron, C.C. Brinton, and P. Gemski

Subjects
Genetics, Salmonella, Circular bacterial chromosome, Biophysics, Chromosome, Cell Biology, Biology, medicine.disease_cause, biology.organism_classification, Biochemistry, Genetic recombination, Pilus, Microbiology, Transduction (genetics), medicine, Molecular Biology, Escherichia coli, Bacteria
Abstract

Pili are protein macromolecules synthesized by bacteria in the form of long, thin, rigid appendages radiating from the cell surface ( Brinton et al. 1954 ). They can be visualized in the electron microscope and are easily concentrated and purified ( Brinton and Stone, 1961 ). Recent observations have indicated that their genetic behavior may be amenable to analysis by the elegant tools of recombination ( Lederberg and Tatum, 1946 ) and transduction ( Zinder and Lederberg, 1952 ). Thus, Brinton and Baron (1960) have reported the transfer of piliation from Escherichia coli to Salmonella typhosa by sexual recombination. The S. typhosa recipient cells gained the ability to produce E. coli pili rather than Salmonella pili, and a possible chromosomal location for the piliation factor close to arabinose on the bacterial chromosome of E. coli was inferred. The transduction of the piliation factor has been demonstrated recently in E. coli by Brinton and Gemski (1961) using phage P1. This paper reports a more precise location of the piliation factor by means of interrupted mating experiments as well as certain findings related to the instability of the piliation factor in E. coli obtained by transductional analysis.

Published
1961

41. Increased survival in calves of Escherichia coli K-12 carrying an Ent plasmid

Author

L P Williams, S Falkow, L D Rollins, and S L Seaman

Subjects
Strain (chemistry), Cell Survival, Immunology, Extrachromosomal Inheritance, Biology, medicine.disease_cause, Microbiology, Extrachromosomal inheritance, Bacterial counts, Infectious Diseases, Plasmid, Escherichia coli, otorhinolaryngologic diseases, medicine, Animals, Cattle, Parasitology, Cell survival, Feces, Plasmids, Research Article
Abstract

Escherichia coli K-12 strains with and without an Ent plasmid were fed to calves, and the survival of each was monitored by viable bacterial counts of the feces. The E. coli K-12 strain carrying the Ent plasmid survived in the calves at significantly higher levels and for a longer period of time than the E. coli F(-) stain.

Published
1976

42. Long-Term Survival in Adult Neuroblastoma with Multiple Recurrences

Author

L. Vénat-Bouvet, V. Le Brun-Ly, J. Martin, O. Gasnier, S. Falkowsky, and N. Tubiana-Mathieu

Subjects
Adults, Long-term survival, Neuroblastoma, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

Neuroblastoma (NB) rarely occurs in adults, and less than 10% of the cases occur in patients older than 10 years. Currently, there are no standard treatment guidelines for adult NB patients. We report the case of a young man suffering from NB in adulthood with multiple recurrences. Treatment included multiple resections, chemotherapy, and radiotherapy. This patient remains free of clinical disease more than 7 years after diagnosis.

Published
2010
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43. Murine model for pertussis vaccine encephalopathy: role of the major histocompatibility complex; antibody to albumin and to Bordetella pertussis and pertussis toxin

Author

L, Steinman, A, Weiss, N, Adelman, M, Lim, J, Oehlert, R, Zuniga, E, Hewlett, and S, Falkow

Subjects
Pertussis Vaccine, Brain Diseases, H-2 Antigens, Immunization, Passive, Mice, Inbred Strains, Serum Albumin, Bovine, Complement System Proteins, Cross Reactions, Bordetella pertussis, Disease Models, Animal, Mice, Pertussis Toxin, Antibody Formation, Mutation, Adenylate Cyclase Toxin, Animals, Virulence Factors, Bordetella
Abstract

A mouse model for pertussis immunization encephalopathy has been described with features that closely resemble the severe adverse reactions occasionally seen after pertussis vaccine administration,m including seizures and a shock-like state leading to death. These reactions are produced with nearly one hundred percent efficiency provided that the mice immunized with Bordetella pertussis have 1) the appropriate major histocompatibility (H-2) genotype, 2) have been sensitized to bovine serum albumin (BSA), and 3) that the injected B. pertussis contained sufficient amounts of pertussis toxin. Antibody titres were measured in mice with haplotypes H-2d.s.k. that are highly susceptible to encephalopathy as well as in H-2b mice, that are totally resistant. Mice with H-2d.s.k. haplotypes were high responders to BSA, while H-2b (B10) mice were non-responders to BSA. Both H-2d and H-2b mice responded well to B. pertussis. Encephalopathy was induced in resistant H-2b mice with B. pertussis and passively administered anti-BSA antiserum, but not with B. pertussis and anti-(T,G)-A--L antibody. This indicated that B. pertussis and anti-BSA were absolutely required for development of encephalopathy. Encephalopathy could be induced in mice decomplemented with cobra venom factor and given BSA and B. pertussis. Several single-site mutants of B. pertussis affecting single virulence factors were induced with transposon Tn5. One of these mutants, BP357, deficient in pertussis toxin production, had a greatly reduced encephalopathic potential in the mouse model compared to the virulent strain BP 338, or to BP348, an adenylate cyclase and hemolysin double mutant, or to BP 349, a hemolysin mutant.(ABSTRACT TRUNCATED AT 250 WORDS)

Published
1985

47. Pilin-gene phase variation of Moraxella bovis is caused by an inversion of the pilin genes

Author

W W Ruehl, S Falkow, Carl F. Marrs, and Gary K. Schoolnik

Subjects
DNA, Bacterial, Inverted repeat, Molecular Sequence Data, Moraxella bovis, Microbiology, Sequence Homology, Nucleic Acid, Gene expression, Coding region, Moraxella, Molecular Biology, Gene, Phase variation, Genetics, Regulation of gene expression, Immunoassay, biology, Base Sequence, Nucleic Acid Hybridization, DNA Restriction Enzymes, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, Molecular biology, Gene Expression Regulation, Genes, Bacterial, Pilin, Fimbriae, Bacterial, Chromosome Inversion, biology.protein, bacteria, Electrophoresis, Polyacrylamide Gel, Fimbriae Proteins, Bacterial Outer Membrane Proteins, Research Article
Abstract

Moraxella bovis Epp63 can express either of two different pilin proteins, called alpha and beta. We have previously cloned and sequenced the beta-pilin gene and now report that DNAs isolated from bacteria expressing alpha pilin have hybridization patterns consistently different from those of bacteria expressing beta pilin. The phase variation between alpha- and beta-pilin gene expression appears to be associated with an inversion of about 2 kilobases of DNA, whose endpoints occur within the coding region of the expressed pilin gene. Comparisons of the beta-pilin gene sequence with those of well-studied bacterial inversion systems revealed a stretch of 58% sequence similarity (21 of 36 base pairs) between the left inverted repeat of the Salmonella typhimurium flagellar hin control region and the amino-terminal portion of the beta-pilin gene.

Published
1988

48. Genetic and molecular characteristics of Vir plasmids of bovine septicemic Escherichia coli

Author

P L Shipley, S Falkow, J Lopez-Alvarez, and C L Gyles

Subjects
Transposable element, DNA, Bacterial, Bacterial Toxins, Bacteriocin Plasmids, Virulence, Cattle Diseases, Biology, medicine.disease_cause, Microbiology, chemistry.chemical_compound, Plasmid, Transformation, Genetic, Sepsis, medicine, Escherichia coli, Animals, Molecular Biology, T-DNA Binary system, Escherichia coli Infections, Plasmid preparation, Antigens, Bacterial, biochemical phenomena, metabolism, and nutrition, chemistry, Conjugation, Genetic, Antigens, Surface, bacteria, Cattle, DNA, Circular, DNA, Plasmids, Research Article
Abstract

Three wild strains of bovine septicemic Escherichia coli were selected on the basis of their production of a toxin lethal for mice and chickens and their characteristic surface antigen. The transfer of these virulence (Vir) properties from two of the three to recipient E. coli was detected after mating. One Vir plasmid (pJL1) was derepressed for transfer and associated with mobilization of chromosomal markers. The other, pJL2, was repressed. Both plasmids were tagged with transposon Tn5 (kanamycin resistance), and transfer parameters of the tagged plasmids were studied. The Tn5 insertion in pJL2 usually increased transfer efficiency 100-fold. Plasmid pJL1 was classified as a member of the FIV incompatibility group. A pJL1::Tn5 derivative plasmid was incompatible with ColV1. Plasmid pJL2 behaved as an fi+ plasmid. Both plasmids pJL1 and pJL2 had a molecular weight of 92 x 10(6) and were present at about four copies per chromosome; their deoxyribonucleic acid (DNA) structures were not identical on the basis of restriction enzyme analysis. DNA-DNA hybridization revealed a polynucleotide sequence homology of at least 58% between the two plasmids. No plasmids could be detected in one wild or certain laboratory-derived Vir+ E. coli strains.

Published
1980

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