Cloning and expression of the Campylobacter jejeuni glya gene in Escherichia coli.

by Ali M.A..* Kibue

Written in English
Published: Pages: 96 Downloads: 946
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  Electroporation permits the uptake of DNA by mammalian cells and plant protoplasts because it induces transient permeability of the cell membrane. We investigated the utility of high-voltage electroporation as a method for genetic transformation of intact bacterial cells by using the enteric pathogen Campylobacter jejuni as a model system. This report demonstrates that the application of . Campylobacter is a major cause of foodborne illnesses worldwide. Campylobacter infections, commonly caused by ingestion of undercooked poultry and meat products, can lead to gastroenteritis and chronic reactive arthritis in humans. Whole genome sequencing (WGS) is a powerful technology that provides comprehensive genetic information about bacteria and is increasingly being applied to study. Campylobacter jejuni and Campylobacter coli are recognized as the most common causative agents of bacterial gastroenteritis in the world. Humans most often become infected by ingesting contaminated food, especially undercooked chicken, but also other sources of bacteria have been described. Campylobacteriosis is normally a self-limiting by: Campylobacter coli and Campylobacter jejuni in chickens Carla M Carvalho1†, Ben W Gannon2†, Deborah E Halfhide2, Silvio B Santos1, Christine M Hayes2, John M Roe2, Joana Azeredo1* Abstract Background: Poultry meat is one of the most important sources of human campylobacteriosis, an acute bacterial enteritis which is a major problem worldwide.

Wang, Fei, "Immunogenic and protective properties of recombinant proteins from a highly pathogenic Campylobacter jejuni clone associated with sheep abortion" (). Campylobacter jejuni clone SA has recently emerged as the prevalent cause of E. coli expression system.   An Escherichia coli (ATCC ) containing a gene encoding a novel maltogenic amylase of Bacillus licheniformis (BLMA), wherein said gene has a length of about kb; said gene expresses a gene product capable of hydrolyzing cyclodextrin, pullulan, as well as starch at an optimum temperature of about 50° C. at a pH of about 7; said gene product has sugar transferase activity in the . Keywords: C. jejuni, C. coli, Propanoate metabolism Background Campylobacter species are a major cause of food-borne disease in the developed and the developing world. Over 80% of human cases are caused by Campylobacter jejuni and around 10% by Campylobacter coli, with the re-maining human cases caused by other Campylobacter species. The Campylobacter jejuni CiaC virulence protein is secreted from the flagellum and delivered to the gene abolishes export of All cloning steps were performed in. E. coli. TOP10 (Invitrogen), and the ACD fusion vectors were transformed into. E. coli.

Campylobacter jejuni strain M1 (laboratory designation 99/) is a rarely documented case of direct transmission of C. jejuni from chicken to a person, resulting in enteritis. We have sequenced the genome of C. jejuni strain M1, and compared this to 12 other C. jejuni sequenced genomes currently publicly available. Compared to these, M1 is closest to strain Bacteria. Campylobacter jejuni F is a hu-man clinical isolate. The cadF gene in F was disrupted by homologous recombination via a single crossover event between the cadF gene on the bac-terial chromosome and an internal fragment of the gene on a suicide vector, as described previously (12). Additional details on methodology may be. commensal and pathogenic behaviours of Campylobacter jejuni. In this study, we provide an example of how second-site mutations can interfere with gene function analysis in C. jejuni. Deletion of the flagellin B gene (flaB)inC. jejuni M1 resulted in mutant clones with inconsistent motility phenotypes.   Analysis of evolutionary patterns of genes in Campylobacter jejuni and C. coli Published on: 08/10/ Author/s: Lars Snipen,1 Trudy Wassenaar,2 Eric Altermann,1,3,10 Jonathan Olson,4 Sophia Kathariou,5 Karin Lagesen,6 Monica Takamiya,7 Susanne Knøchel,7 David W Ussery,8 and Richard J Meinersmann9.

Cloning and expression of the Campylobacter jejeuni glya gene in Escherichia coli. by Ali M.A..* Kibue Download PDF EPUB FB2

Gene, 73 () Eisevier GEN Cloning and expression of the Campy lobacter jejuni glyA gene in Escherichia coli (Recombinant DNA; genomic library; plasmid pBR; serine hydroxymethyltransferase; promoter; comple- mentation) V.L.

Chan, H. Bingham, A. Kibue, P.R.V. Nayudu* and J.L. Penner Department of Microbiology, University of Toronto, Toronto (Canada M5S Cited by: Cloning and expression of the Campylobacter jejuni glyA gene in Escherichia coli.

Gene. Dec 15; 73 (1)– Facklam RR, Padula JF, Thacker LG, Wortham EC, Sconyers BJ. Presumptive identification of group A, B, and D streptococci. Appl Microbiol. Cited by: The basis for the difference between Campylobacter jejuni and Campylobacter coli is the presence and expression of the N-benzoylglycine amidohydrolase (hippuricase) gene only in C.

jejuni. The outer membranes of many gram-negative bacteria contain a major heat-modifiable protein which shows serological cross-reactivity with the OmpA protein of Escherichia coli K Using the cloned gene for the E.

coli K12 protein as a DNA-DNA hybridization Cited by: Abstract. Flagella are essential for motility and have been implicated to be one of the pathogenic determinants. The flagellum ofCampylobacter jejuni is a polymeric structure of a kd protein.

Using a high-affinity flagellin antibody to screen a lambda gt 11 phage genomic expression library ofC. jejuni strain TGH (Serotype LIO36), a recombinant phage clone lambda gt 11RK that expresses Cited by:   The erythromycin resistance gene (Emr) from Campylobacter jejuni ABA94 plasmid DNA was cloned into the pUC18 vector and then expressed in Escherichia coli.

The location of the Emr determinant on the chimeric plasmid was determined by restriction endonuclease mapping within a kb EcoRI fragment. This fragment then hybridized to the kb plasmid DNA but not to the or kb Author: R. Son, A. Ansary. A Bacillus cellulase gene coding for carboxymethylcellulase (CMCase) has been cloned in Escherichia coli using pBR as a gene was expressed independently of its orientation in the cloning vector showing enzyme activity 40 times greater than that produced by the original Bacillus high production of CMCase in E.

coli by the foreign gene did not impede growth of the host Cited by: stream of the pepN gene whose deduced amino acid sequence is homologous to that of the di- and tripeptide transport pro-tein of Lactococcus lactis (6).

In this article, we report the cloning of this gene, the functional expression of the protein in Escherichia coli, and the characterization of this protein as a di-and tripeptide by: Microbiology (1 ),Printed in Great Britain Expression of Campylobacter hyoilei lipo-oligosaccharide (LOS) antigens in Escherichia coli Victoria Korolik,’ Ben N.

Fry,2 Malcolm. High-Throughput Cloning of Campylobacter jejuni ORFs by in Vivo Recombination in Escherichia coli Here, we describe the generation of a Campylobacter jejuni expression clone set using a high-throughput cloning approach based on recombination in E.

coli. The. Cloning and expression of α-Amylase gene from Bacillus subtilis in Pichia pastoris and Escherichia coli. Introduction Enzyme is a type of catalyst that present in living organisms used for many biotechnological functions in various industrial processing.

Campylobacter jejuni, from the delta-epsilon group of proteobacteria, is a microaerophilic, Gram-negative, flagellate, spiral bacterium—properties it shares with the related gastric pathogen.

The thermophilic Campylobacter jejuni and Campylobacter coli are considered weakly clonal populations where incongruences between genetic markers are assumed to be due to random horizontal transfer of genomic DNA. In order to investigate the population genetics structure we extracted a set of core gene families (CGF) from 27 sequenced genomes of C.

jejuni and C. by: 6. The Evolution of Campylobacter jejuni and Campylobacter coli Samuel K. Sheppard1 and Martin C.J. Maiden2 1College of Medicine, Institute of Life Science, Swansea University, Singleton Park, Swansea SA2 8PP, United Kingdom 2Department of Zoology, University of Oxford, Oxford OX1 3PS, United Kingdom Correspondence: @ The global significance of Campylobacter.

Escherichia coli (E. coli) are Gram negative enteric bacteria that live in the human gut. Its presence in nature is an indication of human feacal pollution. coli also causes human disease such as urinary tract infections and neonatal meningitis.

Comparative Gene Expression Analyses of Campylobacter jejuni Strains Isolated from Clinical, Environmental and Animal Sources Ghiwa Azzi Thesis submitted to the Faculty of Graduate and Postdoctoral Studies In partial fulfilment of the requirements for the Escherichia coli strains.

The common antigen components extracted from the isolated C. jejuni strain were examined by SDS-PAGE and Western blot analysis. The 4A7 MAb recognized a protein band of about 62 kDa.

The genomic DNA of C. jejuni was partially digested with EcoRI and HindIII restriction endonuclease. Selected DNA fragments of kb were isolated from agarose gels and ligated into the EcoRI and HindIII Author: Han WenYu, Jiang WenZheng, Lei LianCheng, Yin Zhen. Plasmid DNA was isolated from a positive E.

coli clone. The DNA insert was sequenced and found to contain two open reading frames, one of which encoded a novel esterase (estA). Using different primers the gene was amplified by polymerase chain reaction and inserted into an inducible expression vector (pKK 3) for expression in E.

coli. Abstract. A method for high-resolution genomic fingerprinting of the enteric pathogens Campylobacter jejuni and Campylobacter coli, based on the determination of amplified fragment length polymorphism, is potential of this method for molecular epidemiological studies of these species is evaluated with 50 type, reference, and well-characterised field by:   Campylobacter jejuni infections are a leading cause of bacterial food-borne diarrhoeal illness worldwide, and Campylobacter infections in children Cited by:   Gene encoding an outer membrane protein found in Campylobacter coli and Campylobacter jejuni isolates and methods for detecting virulent Campylobacter spp.

BACKGROUND OF THE INVENTION. This work was supported in part by a grant from the National Institute of Health of the United States government (1R01 DKA1). Abstract. The number of human cases of enteritis caused by Campylobacter jejuni and C.

coli is increasing in Denmark and other European countries. No systematic typing has earlier been performed on Campylobacter isolates of Danish origin. The primary purpose of this study was to provide a serotype distribution of Campylobacter isolates from Danish patients and the major food production Cited by: Molecular cloning and expression in E.

coli of a Salmonella typhi porin gene Isabel Zaror, Isabel G6mez, Gonzalo Castillo, Arturo Yudelevich and Alejandro Venegas Laboratorio de Bioquimica, Pontificia Universidad Catblica de Chile, Casilla D, Santiago, Chile Received 24 December Key words:Immunogenic protein, Campylobacter coli, subunit vaccine Campylobacter spp., gram-negative bacteria, is now the most comm only isolated human enteropathogen.

Human Campylobacter infection predominantly manifests as acute, often bloody diarrhoea accompanied by a fever and abdominal pain (Ketley, ). IN VITRO SURVIVAL OF CAMPYLOBACTER JEJUNI AND CAMPYLOBACTER COLI AT LOW PH Except where reference is made to the work of others, the work described in this thesis is my own or was done in collaboration with my advisory committee.

Campylobacter is currently a leading bacterial cause for human diarrheal illness in the United States, and is, in addition, associated with severe autoimmune sequelae such as Guillain-Barre Syndrome. Poultry are a leading vehicle for human infections, and other food production animals are frequently colonized as well.

However, the ecology of Campylobacter in the farm environment and in its. The survival of Campylobacter jejuni and Campylobacter coli at pH, and for up to 24 h, and the induction of an acid adaptation response in 12 C.

jejuni and 10 C. coli strains in early exponential or late stationary phases in tryptic soy broth (TSB) and Brucella broth were investi-gated. coli strains were more sensitive than. Rise in human infections from Campylobacter and E. coli, whilst Salmonella cases continue to fall: EFSA and ECDC zoonoses report Campylobacteriosis remains the most reported zoonotic disease in humans [1], with a continuous increase in reported cases over the last five years.

Campylobacter jejuni is the leading bacterial cause of human gastroenteritis in the developed world. To improve our understanding of this important human pathogen, the C.

jejuni NCTC genome was sequenced and published in The original annotation was a milestone in Campylobacter research, but is outdated. We now describe the complete re-annotation and re Cited by: gene (fig. 1 and 2). Because of strong conservation of regions in the rRNA genes (5S, 16S and 23S rRNA), these genes are suitable targets for PCR based identification of Campylobacter.

The hippuricase gene (hipO) is specific for C. jejuni and it is not detected in any other Campylobacter species (Kolackova and Karpiskova, ). The hisB gene, found in the enterobacteria (such as E. coli), in Campylobacter jejuni and in Xylella/Xanthomonas encodes a protein involved in catalysis of two step in histidine biosynthesis (the sixth and eight step), In E.

coli hisB is found on the hisGDCBHAFI operon.Internationally recognised Campylobacter experts critically review the most important aspects of Campylobacter research, providing the first coherent picture of the organism's molecular and cellular biology since the publication of the genome. Most contributions are written from a molecular and genomic perspective and contain speculative models upon which to base future research efforts.Antigenicity of the Campylobacter coli CjaA Protein Produced by Escherichia coli ANNA RACZKO, AGNIESZKA WYSZYŃSKA and ELŻBIETA K.

JAGUSZTYN-KRYNICKA* Department of Bacterial Genetics, Institute of Microbiology, Warsaw University, Miecznikowa 1 str. Warsaw, Poland Received 13 November Abstract.