Identification of Escherichia coli flagellar types by restrictionof the amplified fliC gene

2000 ◽  
Vol 151 (7) ◽  
pp. 535-546 ◽  
Author(s):  
Jorge Machado ◽  
Francine Grimont ◽  
Patrick A.D. Grimont
Keyword(s):  
Escherichia Coli ◽  
Flic Gene

scholarly journals GadE regulates fliC gene transcription and motility in Escherichia coli

2020 ◽  
Vol 10 (1) ◽  
pp. 14-23
Author(s):  
William R Schwan ◽  
Nicole L Flohr ◽  
Abigail R Multerer ◽  
Jordan C Starkey
Keyword(s):  
Escherichia Coli ◽  
Gene Transcription ◽  
Flic Gene

scholarly journals Clonal study of avian Escherichia coli strains by fliC conserved-DNA-sequence regions analysis

2008 ◽  
Vol 28 (10) ◽  
pp. 508-514 ◽  
Author(s):  
Tatiana Amabile de Campos ◽  
Gerson Nakazato ◽  
Eliana Guedes Stehling ◽  
Marcelo Brocchi ◽  
Wanderley Dias da Silveira
Keyword(s):  
Escherichia Coli ◽  
Comparative Analysis ◽  
Dna Sequencing ◽  
Dna Sequence ◽  
Yersinia Enterocolitica ◽  
Proteus Mirabilis ◽  
Great Similarity ◽  
Flic Gene ◽  
E Coli ◽  
Zoonotic Risk

The clonal relationship among avian Escherichia coli strains and their genetic proximity with human pathogenic E. coli, Salmonela enterica, Yersinia enterocolitica and Proteus mirabilis, was determined by the DNA sequencing of the conserved 5' and 3'regions fliC gene (flagellin encoded gene). Among 30 commensal avian E. coli strains and 49 pathogenic avian E. coli strains (APEC), 24 commensal and 39 APEC strains harbored fliC gene with fragments size varying from 670bp to 1,900bp. The comparative analysis of these regions allowed the construction of a dendrogram of similarity possessing two main clusters: one compounded mainly by APEC strains and by H-antigens from human E. coli, and another one compounded by commensal avian E. coli strains, S. enterica, and by other H-antigens from human E. coli. Overall, this work demonstrated that fliC conserved regions may be associated with pathogenic clones of APEC strains, and also shows a great similarity among APEC and H-antigens of E. coli strains isolated from humans. These data, can add evidence that APEC strains can exhibit a zoonotic risk.

scholarly journals Identification of new flagellin-encoding fliC genes in Escherichia coli isolated from domestic animals using RFLP-PCR and sequencing methods

2013 ◽  
Vol 33 (4) ◽  
pp. 417-422 ◽  
Author(s):  
Cláudia de Moura ◽  
Monique Ribeiro Tiba ◽  
Marcio José da Silva ◽  
Domingos da Silva Leite
Keyword(s):  
Escherichia Coli ◽  
Fragment Length Polymorphism ◽  
Domestic Animals ◽  
Flic Gene ◽  
E Coli ◽  
New Genes ◽  
Pcr Rflp ◽  
Specific Antisera ◽  
Flagellar Antigen ◽  
Restriction Fragment Length

Identification of Escherichia coli requires knowledge regarding the prevalent serotypes and virulence factors profiles allows the classification in pathogenic/non-pathogenic. However, some of these bacteria do not express flagellar antigen invitro. In this case the PCR-restriction fragment length polymorphism (RFLP-PCR) and sequencing of the fliC may be suitable for the identification of antigens by replacing the traditional serology. We studied 17 samples of E. coli isolated from animals and presenting antigen H nontypeable (HNT). The H antigens were characterized by PCR-RFLP and sequencing of fliC gene. Three new flagellin genes were identified, for which specific antisera were obtained. The PCR-RFLP was shown to be faster than the serotyping H antigen in E. coli, provided information on some characteristics of these antigens and indicated the presence of new genes fliC.

scholarly journals New Approaches for Escherichia coli Genotyping

Pathogens ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 73
Author(s):  
Roman Kotłowski ◽  
Katarzyna Grecka ◽  
Barbara Kot ◽  
Piotr Szweda
Keyword(s):  
Escherichia Coli ◽  
In Silico ◽  
In Silico Analysis ◽  
Point Of View ◽  
Discriminatory Power ◽  
Gene Encoding ◽  
High Genetic Diversity ◽  
Flic Gene ◽  
E Coli ◽  
Complete Sequences

Easy-to-perform, fast, and inexpensive methods of differentiation of Escherichia coli strains beyond the species level are highly required. Herein two new, original tools for genotyping of E. coli isolates are proposed. The first of the developed method, a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) test uses a highly variable fliC gene, encoding the H antigen as a molecular target. The designing of universal pair of primers and selection of the optimal restriction enzyme RsaI was preceded by in silico comparative analysis of the sequences of the genes coding for 53 different serotypes of H-antigen (E. coli flagellin). The target fragments of E. coli genomes for MLST method were selected on the basis of bioinformatics analysis of complete sequences of 16 genomes of E. coli. Initially, seven molecular targets were proposed (seven pairs of primers) and five of them were found useful for effective genotyping of E. coli strains. Both developed methods revealed high differentiation power, and a high genetic diversity of the strains tested was observed. Within the group of 71 strains tested, 29 and 47 clusters were revealed with fliC RFLP-PCR and MLST methods, respectively. Differentiation of the strains with the reference BOX-PCR method revealed 31 different genotypes. The in silico analysis revealed that the discriminatory power of the new MLST method is comparable to the Pasteur and Achtman schemes and is higher than the discriminatory power of the method developed by Clermont. From the epidemiology point of view, the outcomes of our investigation revealed that in most cases, the patients were infected with unique strains, probably from environmental sources. However, some strains isolated from different patients of the wards of pediatrics, internal medicine, and neurology were classified to the same genotype when the results of all three methods were taken into account. It could suggest that they were transferred between the patients.

scholarly journals Cloning, expression and purification of the recombinant FliC from Salmonella enteritidis

2016 ◽  
Vol 19 (4) ◽  
pp. 62-69
Author(s):  
Chau Thi Bao Tran ◽  
Anh Viet Nguyen ◽  
Hieu Van Tran
Keyword(s):  
Escherichia Coli ◽  
Salmonella Enteritidis ◽  
Recombinant Vaccines ◽  
The Novel ◽  
Expression And Purification ◽  
Material Source ◽  
Flic Gene ◽  
Recombinant Vector ◽  
Sds Page ◽  
Adjuvant Property

FliC protein from Salmonella enteritidis is currently interested due to its immunologic adjuvant property for the novel generation of recombinant vaccines. To produce a source for further researches on the immune effects of FliC, we generated an Escherichia coli based on recombinant vector called pET-fliC which is ligated from fliC gene with NdeI and XhoI double digested pET vectors. The results of expression of recombinant FliC, which was induced by IPTG, were confirmed by SDS-PAGE and Western blot probed with anti-6xHis tag. With the purity above 95 %, this recombinant FliC can be used as a material source for next studies on evaluating the adjuvant property of FliC.

Structural organization of escherichia coli ribosomes as determined by immuno electron microscopy

1978 ◽  
Vol 36 (2) ◽  
pp. 166-167 ◽  
Author(s):  
G. Stöffler ◽  
R.W. Bald ◽  
J. Dieckhoff ◽  
H. Eckhard ◽  
R. Lührmann ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Escherichia Coli ◽  
Ribosomal Proteins ◽  
Structural Organization ◽  
Three Dimensional ◽  
Ribosomal Subunit ◽  
Spatial Arrangement ◽  
Small Subunit ◽  
Antibody Binding ◽  
Immuno Electron Microscopy

A central step towards an understanding of the structure and function of the Escherichia coli ribosome, a large multicomponent assembly, is the elucidation of the spatial arrangement of its 54 proteins and its three rRNA molecules. The structural organization of ribosomal components has been investigated by a number of experimental approaches. Specific antibodies directed against each of the 54 ribosomal proteins of Escherichia coli have been performed to examine antibody-subunit complexes by electron microscopy. The position of the bound antibody, specific for a particular protein, can be determined; it indicates the location of the corresponding protein on the ribosomal surface.The three-dimensional distribution of each of the 21 small subunit proteins on the ribosomal surface has been determined by immuno electron microscopy: the 21 proteins have been found exposed with altogether 43 antibody binding sites. Each one of 12 proteins showed antibody binding at remote positions on the subunit surface, indicating highly extended conformations of the proteins concerned within the 30S ribosomal subunit; the remaining proteins are, however, not necessarily globular in shape (Fig. 1).

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