scholarly journals Ectopic Expression of O Antigen in Bordetella pertussis by a Novel Genomic Integration System

mSphere ◽  
2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Keisuke Ishigaki ◽  
Naoaki Shinzawa ◽  
Sayaka Nishikawa ◽  
Koichiro Suzuki ◽  
Aya Fukui-Miyazaki ◽  
...  
Keyword(s):  
Genetic Locus ◽  
Ectopic Expression ◽  
Artificial Chromosome ◽  
Bordetella Bronchiseptica ◽  
Integration System ◽  
O Antigen ◽  
Bacterial Artificial Chromosome Vector ◽  
Number Of Genes ◽  
First Time ◽  
Number Of Bacteria

Some bacterial phenotypes emerge through the cooperative functions of a number of genes residing within a large genetic locus. To transfer the phenotype of one bacterium to another, a means to introduce the large genetic locus into the recipient bacterium is needed. Therefore, we developed a novel system by combining the advantages of a bacterial artificial chromosome vector and phage-derived gene integration machinery. In this study, we succeeded for the first time in introducing a gene locus involved in O antigen biosynthesis ofBordetella bronchisepticainto the chromosome ofB. pertussis, which intrinsically lacks O antigen, and using this system we analyzed phenotypic alterations in the resultant mutant strain ofB. pertussis. The present results demonstrate that this system successfully accomplished the above-described purpose. We consider this system to be applicable to a number of bacteria other thanBordetella.

scholarly journals Genetic Basis for Lipopolysaccharide O-Antigen Biosynthesis in Bordetellae

1999 ◽  
Vol 67 (8) ◽  
pp. 3763-3767 ◽  
Author(s):  
Andrew Preston ◽  
Andrew G. Allen ◽  
Joanna Cadisch ◽  
Richard Thomas ◽  
Kim Stevens ◽  
...  
Keyword(s):  
Insertion Sequence ◽  
Genetic Basis ◽  
Genetic Locus ◽  
Bordetella Bronchiseptica ◽  
Open Reading Frames ◽  
Bordetella Parapertussis ◽  
O Antigen ◽  
Surface Polysaccharide ◽  
Reading Frames

ABSTRACT Bordetella bronchiseptica and Bordetella parapertussis express a surface polysaccharide, attached to a lipopolysaccharide, which has been called O antigen. This structure is absent from Bordetella pertussis. We report the identification of a large genetic locus in B. bronchiseptica and B. parapertussis that is required for O-antigen biosynthesis. The locus is replaced by an insertion sequence in B. pertussis, explaining the lack of O-antigen biosynthesis in this species. The DNA sequence of the B. bronchiseptica locus has been determined and the presence of 21 open reading frames has been revealed. We have ascribed putative functions to many of these open reading frames based on database searches. Mutations in the locus in B. bronchiseptica andB. parapertussis prevent O-antigen biosynthesis and provide tools for the study of the role of O antigen in infections caused by these bacteria.

scholarly journals Ectopic expression of the Stabilin2 gene triggered by an intracisternal A particle (IAP) element in DBA/2J strain of mice

2020 ◽  
Vol 31 (1-2) ◽  
pp. 2-16 ◽  
Author(s):  
Nobuyo Maeda-Smithies ◽  
Sylvia Hiller ◽  
Sharlene Dong ◽  
Hyung-Suk Kim ◽  
Brian J. Bennett ◽  
...  
Keyword(s):  
Promoter Region ◽  
Transmembrane Protein ◽  
Scavenger Receptor ◽  
Ectopic Expression ◽  
Complete Suppression ◽  
Liver Sinusoidal Endothelial Cells ◽  
Normal Expression ◽  
Number Of Genes ◽  
Genes Encoding ◽  
Intracisternal A Particle

AbstractStabilin2 (Stab2) encodes a large transmembrane protein which is predominantly expressed in the liver sinusoidal endothelial cells (LSECs) and functions as a scavenger receptor for various macromolecules including hyaluronans (HA). In DBA/2J mice, plasma HA concentration is ten times higher than in 129S6 or C57BL/6J mice, and this phenotype is genetically linked to the Stab2 locus. Stab2 mRNA in the LSECs was significantly lower in DBA/2J than in 129S6, leading to reduced STAB2 proteins in the DBA/2J LSECs. We found a retrovirus-derived transposable element, intracisternal A particle (IAP), in the promoter region of Stab2DBA which likely interferes with normal expression in the LSECs. In contrast, in other tissues of DBA/2J mice, the IAP drives high ectopic Stab2DBA transcription starting within the 5′ long terminal repeat of IAP in a reverse orientation and continuing through the downstream Stab2DBA. Ectopic transcription requires the Stab2-IAP element but is dominantly suppressed by the presence of loci on 59.7–73.0 Mb of chromosome (Chr) 13 from C57BL/6J, while the same region in 129S6 requires additional loci for complete suppression. Chr13:59.9–73 Mb contains a large number of genes encoding Krüppel-associated box-domain zinc-finger proteins that target transposable elements-derived sequences and repress their expression. Despite the high amount of ectopic Stab2DBA transcript in tissues other than liver, STAB2 protein was undetectable and unlikely to contribute to the plasma HA levels of DBA/2J mice. Nevertheless, the IAP insertion and its effects on the transcription of the downstream Stab2DBA exemplify that stochastic evolutional events could significantly influence susceptibility to complex but common diseases.

scholarly journals Characterization of Bordetella Bronchiseptica Isolated from Rabbits in Fujian, China

2020 ◽  
Author(s):  
Jinxiang Wang ◽  
Shikun Sun ◽  
Yanfeng Chen ◽  
Dongjin Chen ◽  
Lei Sang ◽  
...  
Keyword(s):  
Respiratory Disease ◽  
Animal Species ◽  
Bordetella Bronchiseptica ◽  
Economic Losses ◽  
Fujian Province ◽  
Pcr Screening ◽  
Resistance Rates ◽  
First Time ◽  
Sequence Types

Abstract Background Bordetella bronchiseptica can infect many animal species, and is a potential zoonotic pathogen that can also infect humans. In rabbits, infection of B. bronchiseptica is associated with respiratory disease, which causes economic losses to the rabbit farming. Fujian Province is a traditional importance rabbit farming area in China. However, no literature about the epidemiology and characteristics of B. bronchiseptica in rabbits in Fujian Province has been reported.Results A total of 219 B. bronchiseptica isolates were recovered from the 833 lung samples of dead rabbits with respiratory disease. The 219 isolates were typed into 11 sequence types (STs) including 5 known STs (ST6, ST10, ST12, ST14 and ST33) and 6 new STs (ST88, ST89, ST90, ST91, ST92 and ST93) by using multi-locus sequence typing (MLST). Surprisingly, all the 219 isolates carried the 5 virulence genes of fhaB, prn, cyaA, dnt and bteA in the PCR screening. Moreover, the isolates resistance to cefixime, ceftizoxime, cefatriaxone and ampicillin were detected, and the resistance rates to the 4 kinds of drug were 33.33, 31.05, 11.87 and 3.20%, respectively.Conclusions In the present study, we showed for the first time that B. bronchiseptica is widespread in rabbits in Fujian Province, and that B. bronchiseptica is an important pathogen associating with respiratory disease in rabbits in Fujian Province. Moreover, it should be alert to the potential occurrence of transmission events between rabbits and humans because the B. bronchiseptica strain of ST12 that can infect humans were also isolated from rabbits in Fujian Province.

scholarly journals ASSESSMENT OF THE CURRENT STATE OF COASTAL WATERS OF THE SOUTHERN PART OF LAKE BAIKAL ON SANITARY AND MICROBIOLOGICAL INDICATORS WITH USE OF THE TEST SYSTEMS

2018 ◽  
pp. 54-58
Author(s):  
I.V. Mosharova ◽  
V.V. Il’inskij ◽  
O.V. Il’ina
Keyword(s):  
Lake Baikal ◽  
Coastal Waters ◽  
Test Systems ◽  
Current State ◽  
Wide Range ◽  
Microbial Number ◽  
Southern Site ◽  
First Time ◽  
Number Of Bacteria ◽  
Microbiological Indicators

Sanitary and microbiological researches of a coastal surface water of the southern part of Lake Baikal (from the Listvyanka to the Tanghui) were conducted in June, 2017. Total number of bacteria varied over a wide range – from 0.93 million сells/ml up to 2:05 million сells/ml, with an average 1.41 ± 0.3 million сells/ml. Biomass of bacteria varied from 11.05 to 305.00 mg С/m3, with an average of 123.34 mg С/m3. Coastal waters of the southern site of Lake Baikal mainly had a beta-meso-saprobic status in June, 2017. The total microbial number was determined with the use of the test systems for the first time. The values of the total microbial number were less than 1 000 COU/ml and varied from 9 to 412 COU/ml. Innovative test systems Petrifilm are of great interest for rapid assessments of the sanitary and microbiological status of reservoirs directly in the field conditions.

Dephrin, a transmembrane ephrin with a unique structure, prevents interneuronal axons from exiting the Drosophila embryonic CNS

Development ◽  
2002 ◽  
Vol 129 (18) ◽  
pp. 4205-4218 ◽  
Author(s):  
Torsten Bossing ◽  
Andrea H. Brand
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Ectopic Expression ◽  
Axonal Pathfinding ◽  
N Terminus ◽  
Outer Border ◽  
Embryonic Cns ◽  
First Time ◽  
Neuronal Compartments

Ephrin/Eph signalling is crucial for axonal pathfinding in vertebrates and invertebrates. We identified the Drosophila ephrin orthologue, Dephrin, and describe for the first time the role of ephrin/Eph signalling in the embryonic central nervous system (CNS). Dephrin is a transmembrane ephrin with a unique N terminus and an ephrinB-like cytoplasmic tail. Dephrin binds and interacts with DEph, the Drosophila Eph-like receptor, and Dephrin and DEph are confined to different neuronal compartments. Loss of Dephrin or DEph causes the abberant exit of interneuronal axons from the CNS, whereas ectopic expression of Dephrin halts axonal growth. We propose that the longitudinal tracts in the Drosophila CNS are moulded by a repulsive outer border of Dephrin expression.

Iron-related transcriptomic variations in CaCo-2 cells, an in vitro model of intestinal absorptive cells

2006 ◽  
Vol 26 (1) ◽  
pp. 55-67 ◽  
Author(s):  
Céline Chicault ◽  
Bertrand Toutain ◽  
Annabelle Monnier ◽  
Marc Aubry ◽  
Patricia Fergelot ◽  
...  
Keyword(s):  
Gene Expression ◽  
Functional Annotation ◽  
In Vitro Model ◽  
Iron Absorption ◽  
Intracellular Iron ◽  
Absorptive Cells ◽  
Number Of Genes ◽  
Vitro Model ◽  
First Time

Regulation of iron absorption by duodenal enterocytes is essential for the maintenance of homeostasis by preventing iron deficiency or overload. Despite the identification of a number of genes implicated in iron absorption and its regulation, it is likely that further factors remain to be identified. For that purpose, we used a global transcriptomic approach, using the CaCo-2 cell line as an in vitro model of intestinal absorptive cells. Pangenomic screening for variations in gene expression correlating with intracellular iron content allowed us to identify 171 genes. One hundred nine of these genes are clustered into five types of expression profile. This is the first time that most of these genes have been associated with iron metabolism. Functional annotation of these five clusters indicates potential links between the immune response, proteolysis processes, and iron depletion. In contrast, iron overload is associated with cellular metabolism, especially that of lipids and glutathione involving redox function and electron transfer.

scholarly journals Fluorescence In Situ Hybridization (FISH) on Human Chromosomes Using Photoprobe Biotin-labeled Probes

2003 ◽  
Vol 51 (4) ◽  
pp. 549-551 ◽  
Author(s):  
Anja Weise ◽  
Peter Harbarth ◽  
Uwe Claussen ◽  
Thomas Liehr
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Yeast Artificial Chromosome ◽  
Artificial Chromosome ◽  
Dna Probes ◽  
Human Chromosomes ◽  
First Time ◽  
Established Technique ◽  
Whole Chromosome Painting

Fluorescence in situ hybridization (FISH) on human chromosomes in meta-and interphase is a well-established technique in clinical and tumor cytogenetics and for studies of evolution and interphase architecture. Many different protocols for labeling the DNA probes used for FISH have been published. Here we describe for the first time the successful use of Photoprobe biotin-labeled DNA probes in FISH experiments. Yeast artificial chromosome (YAC) and whole chromosome painting (wcp) probes were tested.

scholarly journals A Single Banana Streak Virus Integration Event in the Banana Genome as the Origin of Infectious Endogenous Pararetrovirus

2008 ◽  
Vol 82 (13) ◽  
pp. 6697-6710 ◽  
Author(s):  
Philippe Gayral ◽  
Juan-Carlos Noa-Carrazana ◽  
Magali Lescot ◽  
Fabrice Lheureux ◽  
Benham E. L. Lockhart ◽  
...  
Keyword(s):  
Artificial Chromosome ◽  
Streak Virus ◽  
Banana Streak Virus ◽  
Genetic Cross ◽  
In The Wild ◽  
Virus Integration ◽  
High Degree ◽  
First Time ◽  
Viral Sequences ◽  
Integration Event

ABSTRACTSequencing of plant nuclear genomes reveals the widespread presence of integrated viral sequences known as endogenous pararetroviruses (EPRVs). Banana is one of the three plant species known to harbor infectious EPRVs.Musa balbisianacarries integrated copies ofBanana streak virus(BSV), which are infectious by releasing virions in interspecific hybrids. Here, we analyze the organization of the EPRV of BSV Goldfinger (BSGfV) present in the wild diploidM. balbisianacv. Pisang Klutuk Wulung (PKW) revealed by the study ofMusabacterial artificial chromosome resources and interspecific genetic cross. cv. PKW contains two similar EPRVs of BSGfV. Genotyping of these integrants and studies of their segregation pattern show an allelic insertion. Despite the fact that integrated BSGfV has undergone extensive rearrangement, both EPRVs contain the full-length viral genome. The high degree of sequence conservation between the integrated and episomal form of the virus indicates a recent integration event; however, only one allele is infectious. Analysis of BSGfV EPRV segregation among an F1 population from an interspecific genetic cross revealed that these EPRV sequences correspond to two alleles originating from a single integration event. We describe here for the first time the full genomic and genetic organization of the two EPRVs of BSGfV present in cv. PKW in response to the challenge facing both scientists and breeders to identify and generate genetic resources free from BSV. We discuss the consequences of this unique host-pathogen interaction in terms of genetic and genomic plant defenses versus strategies of infectious BSGfV EPRVs.

scholarly journals Interactions of Pulmonary Collectins with Bordetella bronchiseptica and Bordetella pertussis Lipopolysaccharide Elucidate the Structural Basis of Their Antimicrobial Activities

2004 ◽  
Vol 72 (12) ◽  
pp. 7124-7130 ◽  
Author(s):  
Lyndsay M. Schaeffer ◽  
Francis X. McCormack ◽  
Huixing Wu ◽  
Alison A. Weiss
Keyword(s):  
Bordetella Pertussis ◽  
Lipid A ◽  
Antimicrobial Activities ◽  
Innate Immune ◽  
Bordetella Bronchiseptica ◽  
Structural Basis ◽  
Wild Type ◽  
Core Oligosaccharide ◽  
The Core ◽  
O Antigen

ABSTRACT Surfactant proteins A (SP-A) and D (SP-D) play an important role in the innate immune defenses of the respiratory tract. SP-A binds to the lipid A region of lipopolysaccharide (LPS), and SP-D binds to the core oligosaccharide region. Both proteins induce aggregation, act as opsonins for neutrophils and macrophages, and have direct antimicrobial activity. Bordetella pertussis LPS has a branched core structure and a nonrepeating terminal trisaccharide. Bordetella bronchiseptica LPS has the same structure, but lipid A is palmitoylated and there is a repeating O-antigen polysaccharide. The ability of SP-A and SP-D to agglutinate and permeabilize wild-type and LPS mutants of B. pertussis and B. bronchiseptica was examined. Previously, wild-type B. pertussis was shown to resist the effects of SP-A; however, LPS mutants lacking the terminal trisaccharide were susceptible to SP-A. In this study, SP-A was found to aggregate and permeabilize a B. bronchiseptica mutant lacking the terminal trisaccharide, while wild-type B. bronchiseptica and mutants lacking only the palmitoyl transferase or O antigen were resistant to SP-A. Wild-type B. pertussis and B. bronchiseptica were both resistant to SP-D; however, LPS mutants of either strain lacking the terminal trisaccharide were aggregated and permeabilized by SP-D. We conclude that the terminal trisaccharide protects Bordetella species from the bactericidal functions of SP-A and SP-D. The O antigen and palmitoylated lipid A of B. bronchiseptica play no role in this resistance.

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